RESUMO
Physiological studies can help predict effects of climate change through determining which species currently live closest to their upper thermal tolerance limits, which physiological systems set these limits, and how species differ in acclimatization capacities for modifying their thermal tolerances. Reductionist studies at the molecular level can contribute to this analysis by revealing how much change in sequence is needed to adapt proteins to warmer temperatures--thus providing insights into potential rates of adaptive evolution--and determining how the contents of genomes--protein-coding genes and gene regulatory mechanisms--influence capacities for adapting to acute and long-term increases in temperature. Studies of congeneric invertebrates from thermally stressful rocky intertidal habitats have shown that warm-adapted congeners are most susceptible to local extinctions because their acute upper thermal limits (LT(50) values) lie near current thermal maxima and their abilities to increase thermal tolerance through acclimation are limited. Collapse of cardiac function may underlie acute and longer-term thermal limits. Local extinctions from heat death may be offset by in-migration of genetically warm-adapted conspecifics from mid-latitude 'hot spots', where midday low tides in summer select for heat tolerance. A single amino acid replacement is sufficient to adapt a protein to a new thermal range. More challenging to adaptive evolution are lesions in genomes of stenotherms like Antarctic marine ectotherms, which have lost protein-coding genes and gene regulatory mechanisms needed for coping with rising temperature. These extreme stenotherms, along with warm-adapted eurytherms living near their thermal limits, may be the major 'losers' from climate change.
Assuntos
Adaptação Fisiológica/fisiologia , Mudança Climática , Adaptação Fisiológica/genética , Animais , Evolução Biológica , Temperatura Corporal/fisiologia , Regulação da Temperatura Corporal/fisiologia , DNA/metabolismo , Malato Desidrogenase/química , Modelos Moleculares , Polimorfismo Genético , Conformação Proteica , TemperaturaRESUMO
The muscle-type (M4) lactate dehydrogenases of Sebastolobus altivelis, a deep-water scorpaenid, and S. alascanus, a shallower species, are electrophoretically indistinguishable, yet differ in pressure sensitivities. The lactate dehydrogenase of S. altivelis exhibits lower pressure sensitivities of substrate and coenzyme binding and catalytic rate. Such apparently pressure-adaptive kinetic properties may be important for establishing species depth zonation patterns in the ocean.
Assuntos
Adaptação Biológica , Peixes/fisiologia , Pressão Hidrostática , L-Lactato Desidrogenase/metabolismo , Pressão , Animais , Evolução Biológica , Isoenzimas , Cinética , Músculos/enzimologia , NAD/metabolismo , Piruvatos/metabolismo , Especificidade da Espécie , TemperaturaRESUMO
Solemya reidi, a gutless clam found in sulfide-rich habitats, contains within its gills bacterial symbionts thought to oxidize sulfur compounds and provide a reduced carbon food source to the clam. However, the initial step or steps in sulfide oxidation occur in the animal tissue, and mitochondria isolated from both gill and symbiont-free foot tissue of the clam coupled the oxidation of sulfide to oxidative phosphorylation [adenosine triphosphate (ATP) synthesis]. The ability of Solmya reidi to exploit directly the energy in sulfide for ATP synthesis is unprecedented, and suggests that sulfide-habitat animals that lack bacterial symbionts may also use sulfide as an inorganic energy source.
RESUMO
Three species of Antarctic fishes which live in constantly near-freezing waters have a markedly low upper-lethal temperature of 6 degrees C ; this is the lowest upper-lethal temperature reported for any organism. The fishes survive supercooling to -2.5 degrees C. Data on brain metabolism in vitro support the hypothesis that the central nervous system is a primary site of thermal injury.
Assuntos
Temperatura Baixa/efeitos adversos , Peixes/fisiologia , Adaptação Fisiológica , Animais , Regiões Antárticas , Encéfalo/metabolismo , Consumo de OxigênioRESUMO
Respiration of plume tissue of the hydrothermal vent tube worm Riftia pachyptila is insensitive to sulfide poisoning in contrast to tissues of animals that do not inhabit vents. Permeability barriers may not be responsible for this insensitivity since plume homogenates are also resistant to sulfide poisoning. Cytochrome c oxidase of plume, however, is strongly inhibited by sulfide at concentrations less than 10 microM. Factors present in blood, but not in cytosol, prevent sulfide from inhibiting cytochrome c oxidase. Avoidance of sulfide poisoning of respiration in Riftia pachyptila thus appears to involve a blood-borne factor having a higher sulfide affinity than that of cytochrome c oxidase, with the result that appreciable amounts of free sulfide are prevented from accumulating in the blood and entering the intracellular compartment.
RESUMO
Striking convergent evolution is found in the properties of the organic osmotic solute (osmolyte) systems observed in bacteria, plants, and animals. Polyhydric alcohols, free amino acids and their derivatives, and combinations of urea and methylamines are the three types of osmolyte systems found in all water-stressed organisms except the halobacteria. The selective advantages of the organic osmolyte systems are, first, a compatibility with macromolecular structure and function at high or variable (or both) osmolyte concentrations, and, second, greatly reduced needs for modifying proteins to function in concentrated intracellular solutions. Osmolyte compatibility is proposed to result from the absence of osmolyte interactions with substrates and cofactors, and the nonperturbing or favorable effects of osmolytes on macromolecular-solvent interactions.
Assuntos
Evolução Biológica , Equilíbrio Hidroeletrolítico , Aminoácidos/fisiologia , Animais , Transporte Biológico Ativo , Glicerol/fisiologia , Íons/fisiologia , Metilaminas/fisiologia , Conformação Molecular , Ureia/fisiologia , Água/fisiologiaRESUMO
1. Comparisons of homologous enzymes from species adapted to widely different temperatures reveal that ligand-binding affinities are rigorously conserved. This is interpreted to mean that a critical relationship between ligand-binding ability and intracellular ligand concentrations must be maintained for proper enzymic regulation. 2. The catalytic efficiencies of enzyme homologues differ in temperature-compensatory manners. Activation free energies are proportional to adaptation temperature and, consequently, low-temperature-adapted enzymes have the highest substrate turnover numbers. 3. Temperature compensatory adjustments in catalytic efficiency may be achieved by altering the number of weak bonds that form or break during a catalytic conformational change. Support for this hypothesis comes from the finding that activation enthalpy and activation entropy values co-vary in a regular manner and by magnitudes consistent with different amounts of weak-bond formation/rupture during catalytic activation in differnt enzyme homologues. 4. Adaptive adjustments in ligand-binding energetics may also involve utilization of the energy changes that occur during conformational changes. This mechanism would permit enzymes with identical binding-site chemistries to display adaptively different ligand affinities. 5. The greater heat-stabilities of enzymes from warm-adapted species may cause these enzymes to be less efficient catalysts than cold-adapted heat-labile enzymes. Heat-stable enzymes may have to break more weak bonds during a catalytic conformational change than do cold-adapted enzymes. The requirements for thermal stability and high catalytic efficiency thus appear to force an adaptational 'compromise'.
Assuntos
Adaptação Fisiológica , Evolução Biológica , Enzimas/metabolismo , Temperatura , Animais , Sítios de Ligação , Peixes , Isoenzimas/metabolismo , Cinética , Conformação Proteica , Piruvato Quinase/metabolismo , Coelhos , Especificidade da EspécieRESUMO
In white skeletal muscle of conspecific pelagic fishes, the activity of enzymes associated with anaerobic glycolysis, e.g., lactate dehydrogenase (LDH), usually scale positively with increasing body size; this pattern is opposite to that found for enzymes of aerobic metabolism, which decrease in mass-specific activity with size (1-3). The higher mass-specific capacities for anaerobic ATP generation in larger conspecifics are thought to facilitate conservation of high-speed ("burst") swimming ability in fishes of different sizes (1). To investigate the mechanisms responsible for scaling of LDH activity, total RNA, and the specific mRNA for LDH-A (the skeletal muscle isoform of the enzyme) in white muscle of paralabrax nebulifer, the barred sand bass. We also measured total protein concentration and the concentration of actin, the major protein of thin filaments, and its specific mRNA. Although LDH activity scaled significantly with body size as predicted (1-4), no other biochemical trait measured showed a significant size-dependent concentration. We conclude that the regulation of LDH activity in white muscle of this species is not governed by LDH-A mRNA concentrations, but rather by one or more other mechanisms, for example rate of translation of LDH message or a reduced rate of degradation of LDH-A in larger fish.
Assuntos
Bass/crescimento & desenvolvimento , L-Lactato Desidrogenase/análise , Músculo Esquelético/enzimologia , RNA Mensageiro/análise , Animais , Constituição Corporal , Regulação da Expressão Gênica , Isoenzimas , L-Lactato Desidrogenase/genéticaRESUMO
Marine intertidal organisms are subjected to a variety of abiotic stresses, including aerial exposure and wide ranges of temperature. Intertidal species generally have higher thermal tolerance limits than do subtidal species, and tropical species have higher thermal tolerance limits than do temperate species. The adaptive significance of upper thermal tolerance limits of intertidal organisms, however, has not been examined within a comparative context. Here, we present a comparative analysis of the adaptive significance of upper thermal tolerance limits in 20 congeneric species of porcelain crabs, genus Petrolisthes, from intertidal and subtidal habitats throughout the eastern Pacific. Upper thermal tolerance limits are positively correlated with surface water temperatures and with maximal microhabitat temperatures. Analysis of phylogenetically independent contrasts (from a phylogenetic tree on the basis of the 16s rDNA gene sequence) suggests that upper thermal tolerance limits have evolved in response to maximal microhabitat temperatures. Upper thermal tolerance limits increased during thermal acclimation at elevated temperatures, the amount of increase being greater for subtidal than for intertidal species. This result suggests that the upper thermal tolerance limits of some intertidal species may be near current habitat temperature maxima, and global warming thus may affect the distribution limits of intertidal species to a greater extent than for subtidal species.
Assuntos
Aclimatação/fisiologia , Braquiúros/fisiologia , Temperatura Alta/efeitos adversos , Filogenia , Altitude , Animais , California , Chile , Modelos Lineares , México , Mortalidade , Oregon , Oceano Pacífico , Panamá , Análise de Regressão , Água do Mar , TemperaturaRESUMO
The time course and magnitude of the heat-shock response in relation to severity of thermal stress are important, yet poorly understood, aspects of thermotolerance. We examined patterns of protein synthesis in congeneric marine snails (genus Tegula) that occur at different heights along the subtidal to intertidal gradient after a thermal exposure (30 degrees C for 2.5 h, followed by 50 h recovery at 13 degrees C) that induced the heat-shock response. We monitored the kinetics and magnitudes of protein synthesis by quantifying incorporation of 35S-labeled methionine and cysteine into newly synthesized proteins and observed synthesis of putative heat-shock proteins (hsp's) of size classes 90, 77, 70, and 38 kDa. In the low- to mid-intertidal species, Tegula funebralis, whose body temperature frequently exceeds 30 degrees C during emersion, synthesis of hsp's commenced immediately after heat stress, reached maximal levels 1-3 h into recovery, and returned to prestress levels by 6 h, except for hsp90 (14 h). In contrast, in the low-intertidal to subtidal species, Tegula brunnea, for which 2.5 h at 30 degrees C represents a near lethal heat stress, synthesis of hsp's commenced 2-14 h after heat stress; reached maximal levels after 15-30 h, which exceeded magnitudes of synthesis in T. funebralis; and returned to prestress levels in the case of hsp90 (50 h) and hsp77 (30 h) but not in the case of hsp70 and hsp38. Exposures to 30 degrees C under aerial (emersion) and aquatic (immersion) conditions resulted in differences in hsp synthesis in T. brunnea but not in T. funebralis. The different time courses and magnitudes of hsp synthesis in these congeners suggest that the vertical limits of their distributions may be set in part by thermal stress.
Assuntos
Proteínas de Choque Térmico/biossíntese , Resposta ao Choque Térmico/fisiologia , Caramujos/fisiologia , Animais , Autorradiografia/veterinária , California , Eletroforese em Gel de Poliacrilamida/veterinária , Brânquias/química , Proteínas de Choque Térmico HSP70/biossíntese , Proteínas de Choque Térmico HSP70/química , Proteínas de Choque Térmico HSP90/biossíntese , Proteínas de Choque Térmico HSP90/química , Proteínas de Choque Térmico/química , Temperatura Alta/efeitos adversos , Cinética , Oceano Pacífico , Água do Mar , Caramujos/químicaRESUMO
Most of the important functional and structural properties of enzymes are affected by temperature. In order to maintain critical enzymic properties such as regulatory sensitivity, catalytic potential and structural stability, significant changes have been made in enzymes during evolution in different thermal regimes. Regulatory function, as typified by substrate binding ability, has been especially conservative. For a given enzyme, substrate binding ability is maintained at a relatively stable level over the entire temperature range experienced by the organism (enzyme), in spite of wide variation in substrate affinity at temperatures outside the biological range. Similarities in substrate affinity among homologues and analogues of enzymes from bacteria, invertebrates, fishes and mammals, at respective physiological temperatures for the enzymes, demonstrate the crucial importance of regulatory abilities in enzymes. Two facts, (a) that enzymes function at sub-maximal rates, and (b) that low temperature compensation is not effected by wholesale reductions in apparent Km values, argue that regulation outweighs sheer catalytic potential in enzymic function. The efficiency of an enzyme to catalyze a reaction at a rapid rate appears highest in low cell-temperature forms. The finding that catalytic efficiency is inversely correlated with enzymic heat stability suggests that enzymes with relatively great abilities to undergo conformational changes during catalysis are capable of supplying the most energy for activation events, this energy arising in part from the exergonic formation of weak bonds during the activation step in catalysis. Energy changes due to conformational changes may also be used to reduce the net enthalpy change which occurs during ligand binding, a mechanism we refer to as "coupled-compensating enthalpy changes." Comparisons of amino acid compositions of enzyme homologues and analogues from differently thermally adapted species do not reveal major differences, for example, in the overall hydrophobicity of enzymes. We propose that observed differences in enzyme thermal stability derive more from quantitative differences, i.e., differences in total numbers of secondary interactions, than from quilitative differences, i.e., differences in the relative importance of different classes of weak bonds.
Assuntos
Adaptação Fisiológica , Enzimas/metabolismo , Temperatura , Sequência de Aminoácidos , Animais , Sítios de Ligação , Evolução Biológica , Aves/metabolismo , Ativação Enzimática , Peixes/metabolismo , Humanos , Ligação de Hidrogênio , Cinética , Ligantes , Mamíferos/metabolismo , Conformação Molecular , Oxiemoglobinas/análise , Fosfoenolpiruvato , Piruvato Quinase/metabolismo , Relação Estrutura-AtividadeRESUMO
The composition of the intracellular milieu shows striking similarities among widely different species. Only certain values of intracellular pH, values that generally reflect alphastat regulation, and only narrow ranges of inorganic ion concentrations are found in the cytoplasm of the cells of most animals, plants, and microorganisms. In water-stressed organisms only a few types of low-molecular-weight organic molecules (osmolytes) are accumulated. These highly conserved characteristics of the intracellular fluids reflect the need to maintain critical features of macromolecules within narrow ranges optimal for life. For proteins these features include maintaining adequate rates of catalysis, a high level of regulatory responsiveness, and a precise balance between stability and lability of structure (tertiary conformation, subunit assembly, and multiprotein complexes). The optimal values for these functional and structural features of proteins often lie near the midrange of possible values for these properties, and only under specific conditions of intracellular pH, ionic strength, and osmolyte composition are these optimal midrange values conserved. In dormant cells the departure of solution conditions from values that are optimal for protein function and structure may be instrumental in reducing or shutting down metabolic functions. Seen from a broad evolutionary perspective, the evolution of the intracellular milieu is an important complement to macromolecular evolution. In certain instances appropriate modifications of the internal milieu may reduce the need for adaptive amino acid replacements in proteins.
Assuntos
Líquidos Corporais/fisiologia , Líquido Intracelular/fisiologia , Osmose , Proteínas/fisiologia , Prótons , Equilíbrio Ácido-Base , Adaptação Fisiológica , Aminoácidos/metabolismo , Animais , Evolução Biológica , Temperatura Corporal , Soluções Tampão , Calmodulina/metabolismo , Metabolismo dos Carboidratos , Células/metabolismo , Enzimas/metabolismo , Glicólise , Hibernação , Humanos , Concentração de Íons de Hidrogênio , Substâncias Macromoleculares/fisiologia , Peso Molecular , Equilíbrio HidroeletrolíticoRESUMO
Protein adaptations to the physical environment play critical roles in determining the biogeographical distributions of species. The study of closely related species in habitats differing slightly in temperature or pressure offers an excellent experimental approach for discerning environmental thresholds of protein perturbation and the types of amino acid substitutions that are effective in maintaining optimal protein properties. These adaptations may involve amino acid replacements other than at the active site residues involved in catalysis or binding, a discovery with implications for the debate between adherents to the Neutralist and Selectionist points of view.
RESUMO
Deep-sea ecosystems contain unique endemic species whose distributions show strong vertical patterning in the case of pelagic animals and sharp horizontal patterning in the case of benthic animals living in or near the deep-sea hydothermal vents. This review discusses the biochemical adaptations that enable deep-sea animals to exploit diverse deep-sea habitats and that help establish biogeographic patterning in the deep-sea. The abilities of deep-sea animals to tolerate the pressure and temperature conditions of deep-sea habitats are due to pervasive adaptations at the biochemical level: enzymes exhibit reduced perturbation of function by pressure, membranes have fluidities adapted to deep-sea pressures and temperatures, and proteins show enhanced structural stability relative to homologous proteins from cold-adapted shallow-living species. Animals from the warmest habitable regions of hydrothermal vent ecosystems have enzymes and mitochondria adapted to high pressure and relatively high temperatures. The low metabolic rates of bathypelagic fishes correlate with greatly reduced capacities for ATP turnover in locomotory muscle. Reduced light and food availability in bathypelagic regions select for low rates of energy expenditure in locomotory activity. Deep-sea animals thus reflect the importance of biochemical adaptations in establishing species distribution patterns and appropriate rates of metabolic turnover in different ecosystems.
Assuntos
Adaptação Fisiológica , Bioquímica , Ecologia , Biologia Marinha , Animais , Fenômenos Bioquímicos , Pressão Hidrostática , ATPase Trocadora de Sódio-Potássio/metabolismo , TemperaturaRESUMO
The importance of adaptation to high pressure has long been implicit in the findings of studies in which 1 atm-adapted species were subjected to elevated pressures. Recent comparative studies have shown that pressure sensitivities of enzymes, structural proteins, and membrane-based systems differ markedly between shallow- and deep-living species. These studies allow operational definition of what constitutes high pressures for different biological structures and processes. These are the habitat (adaptation) pressures at which a given type of system first exhibits reduced perturbation by pressure. These threshold pressures vary among physiological systems, but are similar for a given system among different species. Dehydrogenase enzymes and adenylyl cyclases exhibit threshold perturbation pressures of only 50-100 atm; the Na(+)-K(+)-ATPase of teleost gills appears to have a pressure perturbation threshold near 200 atm, and a similar threshold was found for actin self-assembly. Even this limited sample of physiological processes indicates that the terms deep and high pressure begin to apply at depths of only 500 m or less--and processes yet to be examined in comparative analysis may yield even lower pressure thresholds. The differences in sensitivity to pressure of homologous systems in shallow- and deep-living organisms have implications at several levels of biological organization. The vertical distribution patterns of species in aquatic habitats may be established, in part, by interspecific differences in resistance to pressure. High pressures may restrict the depths to which shallow-living species can penetrate, and the obligately barophilic systems found in deep-living organisms may limit their upper distribution limits. The similarities noted among the adaptations of deep-sea species with different shallow-water ancestors reflect a high degree of convergent evolution in pressure adaptation. It will be interesting to learn if the similarities in pressure-resistance of function among diverse deep-sea species are the result of similar or identical changes at the molecular level, e.g. in protein sequence. Acclimation to pressure may be of widespread occurrence among species that undergo large changes in depth, e.g. during ontogeny. Pressure acclimation may require pressure-regulation of gene expression. Lastly, comparisons of species from the cold deep sea with those from hydrothermal vents have shown that adaptations to both temperature and pressure play critical roles in determining the distribution patterns of deep-living species.
Assuntos
Adaptação Fisiológica , Pressão Hidrostática , Animais , Pressão Atmosférica , Estabilidade de Medicamentos , Enzimas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Biossíntese de Proteínas , Receptores Purinérgicos/fisiologia , Transdução de Sinais/fisiologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Transcrição GênicaRESUMO
1. Pyruvate kinase of Alaskan king-crab leg muscle exists in two kinetically distinct forms, each of which displays a different temperature-dependence in the K(m) for phosphoenolpyruvate. 2. A ;cold' variant of the enzyme has hyperbolic kinetics and exhibits a minimal K(m) for substrate at 5 degrees . At physiological concentrations of phosphoenolpyruvate the ;cold' enzyme is active only below 10 degrees . A ;warm' pyruvate kinase has a minimal K(m) for substrate at about 12 degrees . This enzyme displays sigmoidal kinetics and is likely to be inactive, at physiological substrate concentrations, at temperatures below 9 degrees . 3. The combined activities of these two pyruvate kinases yield highly temperature-independent rates of catalysis, at physiological substrate concentrations, over the range of habitat temperatures encountered by the organism, namely 4-12 degrees . 4. The two variants of pyruvate kinase do not appear to be isoenzymes in the conventional sense. Electrophoretic and electrofocus analyses revealed only single peaks of activity. 5. The results suggest that the ;warm' pyruvate kinase and the ;cold' pyruvate kinase are formed by a temperature-dependent interconversion of one protein species. This interconversion has major adaptive significance: as the temperature is lowered the ;warm' enzyme is converted into the ;cold' enzyme; the opposite situation obtains when the temperature is raised. Temperature changes thus mimic the effects noted for fructose 1,6-diphosphate on certain mammalian pyruvate kinases.
Assuntos
Artrópodes/enzimologia , Piruvato Quinase/fisiologia , Animais , Eletroforese , Hexosefosfatos/farmacologia , Cinética , Piruvatos/análise , Piruvatos/metabolismo , TemperaturaRESUMO
Comparative studies of lactate dehydrogenases (LDHs) and skeletal muscle actins from vertebrates adapted to widely different temperatures and hydrostatic pressures reveal major conservative trends in protein evolution and adaptation. For enzymes, ligand binding, as estimated by apparent Michaelis constant (Km) values, is strongly conserved at physiological temperatures, pressures, intracellular pH values and osmotic compositions of different organisms. The catalytic rate constants (kcat values) of enzyme homologues are highest for enzymes of low-body-temperature organisms, a trend that can be interpreted in terms of temperature compensation of metabolism. For skeletal muscle actins, the enthalpy and entropy changes accompanying the assembly of filamentous (F) actin from globular (G) actin are highest in high-body-temperature species and especially low in polar and deep-sea fishes. The thermal stability of G-actin is positively correlated with adaptation temperature, except in the case of actins of deep-sea fishes, which are also highly heat stable. Hydrophobic interactions between actin subunits may be of reduced importance in low-body-temperature animals and, especially, in deep-sea fishes. The differences in enthalpy and entropy changes during the G-to-F transformation favor a close conservation of the equilibrium constant for actin assembly under physiological conditions of temperature and pressure for different species. These adaptive patterns in enzymes and actin are likely to reflect changes in protein primary structure. The appropriate values for protein traits such as ligand binding abilities and catalytic rates are also shown to be established by the composition of the low molecular weight constituents of the cytosol. For example, the use of a combination of urea and methylamine solutes for osmoregulation by marine elasmobranchs is shown to be a mechanism which permits the conservation of key protein traits at high osmolarities. The methylamine solutes such as trimethylamine-N-oxide have effects on proteins opposite to those of urea, and at the approximately 2:1 concentration ratio of urea to methylamines, these counteracting effects are virtually complete. Regulation of hydrogen ion activity (pH) also is shown to play a major role in the conservation of critical protein traits. The importance of temperature-dependent pH in ectotherms is discussed in terms of stabilizing binding abilities and maintaining correct regulatory and structural sensitivities of proteins. The buffering capacity of tissues reflects the potential of the tissue for generating acidic end-products during anaerobic metabolism. Skeletal muscle, especially white locomotory muscle of fishes, is highly buffered relative to red locomotory muscle and heart muscle.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Adaptação Fisiológica , Meio Ambiente , Proteínas/metabolismo , Vertebrados/metabolismo , Actinas/metabolismo , Animais , Concentração de Íons de Hidrogênio , Pressão Hidrostática , Líquido Intracelular/metabolismo , L-Lactato Desidrogenase/metabolismo , Músculos/metabolismo , Concentração Osmolar , Relação Estrutura-Atividade , TemperaturaRESUMO
Changes in enzyme conformation are often accompanied by large changes in volume. Model compound studies suggest that these volume changes may derive from two sources: (i) "hydration density" effects due to changes in the exposure to solvent of protein groups which modify water density, and (ii) "structural" contributions arising from changes in the volume of the protein itself. An experimental approach was developed to test the validity of the predictions based on model compound studies for catalytic conformational changes. By examining the effects of different solutes on the activation volumes of different enzymic reactions, we show that both sources of volume change provide significant contributions to the activation volume.
Assuntos
Ativação Enzimática , Enzimas/metabolismo , Conformação Proteica , Isocitrato Desidrogenase/metabolismo , Cinética , L-Lactato Desidrogenase/metabolismo , Malato Desidrogenase/metabolismo , Matemática , Peso Molecular , Concentração Osmolar , Piruvato Quinase/metabolismo , SoluçõesRESUMO
There exists a linear correlation between the effect of a salt on the rate of an enzymic reaction and its effect on the activation volume (delta V++) of the reaction. Salts that increase delta V++ invariably decrease the rate of the reaction, and vice versa. The salt effects on reaction rate are, however, much larger than would be predicted solely on the basis of pressure-volume work changes deriving from the observed alterations in delta V++. Different inorganic salts affect reaction rates and activation volumes in a manner that reflects the salts' positions in the Hofmeister series. These observations, taken in conjunction with data on the effects of salts on protein functional group (aminoacid side-chains and peptide linkages) hydration, lead us to propose the following hypothesis to account for salt activation and inhibition of catalysis. Aminoacid side-chains and peptide linkages located on or near the protein surface change their exposure to water during conformational events in catalysis. These protein group transfers are accompanied by large volume and energy changes that are due largely to changes in the organization of water around these groups. When these transfer processes occur during the rate-limiting step in catalysis, these energy and volume changes can contribute to the free energy of activation (delta G++) and the activation volume of the reaction. By influencing the degree to which water can organize around transferred protein groups, salts can modify both the delta G++ (rate) and the delta V++ of a reaction.
Assuntos
Enzimas , Íons , Água , Catálise , Fenômenos Químicos , Físico-Química , Enzimas/metabolismo , Isocitrato Desidrogenase/metabolismo , L-Lactato Desidrogenase/metabolismo , Malato Desidrogenase/metabolismo , Concentração Osmolar , Conformação Proteica , Piruvato Quinase/metabolismo , TermodinâmicaRESUMO
Intracellular fluids of marine elasmobranchs (sharks, skates and rays), holocephalans and the coelacanth contain urea at concentrations averaging 0.4m, high enough to significantly affect the structural and functional properties of many proteins. Also present in the cells of these fishes are a family of methylamine compounds, largely trimethylamine N-oxide with some betaine and sarcosine, and certain free amino acids, mainly beta-alanine and taurine, whose total concentration is approx. 0.2m. These methylamine compounds and amino acids have been found to be effective stabilizers of protein structure, and, at a 1:2 molar concentration ratio of these compounds to urea, perturbations of protein structure by urea are largely or fully offset. These counteracting effects of solutes on proteins are seen for: (1) thermal stability of protein secondary and tertiary structure (bovine ribonuclease); (2) the rate and extent of enzyme renaturation after acid denaturation (rabbit and shark lactate dehydrogenases); and (3) the reactivity of thiol groups of an enzyme (bovine glutamate dehydrogenase). Attaining osmotic equilibrium with seawater by these fishes has thus involved the selective accumulation of certain nitrogenous metabolites that individually have significant effects on protein structure, but that have virtually no net effects on proteins when these solutes are present at elasmobranch physiological concentrations. These experiments indicate that evolutionary changes in intracellular solute compositions as well as in protein amino acid sequences can have important roles in intracellular protein function.