RESUMO
The Ramachandran map clearly delineates the regions of accessible conformational (φ-ψ) space for amino acid residues in proteins. Experimental distributions of φ, ψ values in high-resolution protein structures, reveal sparsely populated zones within fully allowed regions and distinct clusters in apparently disallowed regions. Conformational space has been divided into 14 distinct bins. Residues adopting these relatively rare conformations are presented and amino acid propensities for these regions are estimated. Inspection of specific examples in a completely "arid", fully allowed region in the top left quadrant establishes that side-chain and backbone interactions may provide the energetic compensation necessary for populating this region of φ-ψ space. Asn, Asp, and His residues showed the highest propensities in this region. The two distinct clusters in the bottom right quadrant which are formally disallowed on strict steric considerations correspond to the gamma turn (C7 axial) conformation (Bin 12) and the i + 1 position of Type II' ß turns (Bin 13). Of the 516 non-Gly residues in Bin 13, 384 occupied the i + 1 position of Type II' ß turns. Further examination of these turn segments revealed a high propensity to occur at the N-terminus of helices and as a tight turn in ß hairpins. The ß strand-helix motif with the Type II' ß turn as a connecting element was also found in as many as 57 examples.
Assuntos
Aminoácidos/química , Estrutura Molecular , Conformação Proteica , Proteínas/química , Modelos MolecularesRESUMO
Acquired immunodeficiency syndrome (AIDS) is one of the most challenging infectious diseases to treat on a global scale. Understanding the mechanisms underlying the development of drug resistance is necessary for novel therapeutics. HIV subtype C is known to harbor mutations at critical positions of HIV aspartic protease compared to HIV subtype B, which affects the binding affinity. Recently, a novel double-insertion mutation at codon 38 (L38HL) was characterized in HIV subtype C protease, whose effects on the interaction with protease inhibitors are hitherto unknown. In this study, the potential of L38HL double-insertion in HIV subtype C protease to induce a drug resistance phenotype towards the protease inhibitor, Saquinavir (SQV), was probed using various computational techniques, such as molecular dynamics simulations, binding free energy calculations, local conformational changes and principal component analysis. The results indicate that the L38HL mutation exhibits an increase in flexibility at the hinge and flap regions with a decrease in the binding affinity of SQV in comparison with wild-type HIV protease C. Further, we observed a wide opening at the binding site in the L38HL variant due to an alteration in flap dynamics, leading to a decrease in interactions with the binding site of the mutant protease. It is supported by an altered direction of motion of flap residues in the L38HL variant compared with the wild-type. These results provide deep insights into understanding the potential drug resistance phenotype in infected individuals.
Assuntos
Infecções por HIV , Inibidores da Protease de HIV , HIV-1 , Humanos , Saquinavir/química , Saquinavir/farmacologia , Inibidores da Protease de HIV/química , Inibidores da Protease de HIV/farmacologia , HIV-1/genética , Protease de HIV/genética , Farmacorresistência Viral/genéticaRESUMO
Short range side chain-backbone hydrogen bonded motifs involving Asn and Gln residues have been identified from a data set of 1370 protein crystal structures (resolution ≤ 1.5 Å). Hydrogen bonds involving residues i - 5 to i + 5 have been considered. Out of 12,901 Asn residues, 3403 residues (26.4%) participate in such interactions, while out of 10,934 Gln residues, 1780 Gln residues (16.3%) are involved in these motifs. Hydrogen bonded ring sizes (C(n), where n is the number of atoms involved), directionality and internal torsion angles are used to classify motifs. The occurrence of the various motifs in the contexts of protein structure is illustrated. Distinct differences are established between the nature of motifs formed by Asn and Gln residues. For Asn, the most highly populated motifs are the C(10)(CO(δ)(i) NH(i + 2)), C(13)(CO(δ)(i) NH(i + 3)) and C(17)(N(δ)H(i) CO(i - 4)) structures. In contrast, Gln predominantly forms C(16)(CO(ε)(i) NH(i - 3)), C(12)(N(ε)H(i) CO(i - 2)), C(15)(N(ε)H(i) CO(i - 3)) and C(18)(N(ε)H(i) CO(i - 4)) motifs, with only the C(18) motif being analogous to the Asn C(17) structure. Specific conformational types are established for the Asn containing motifs, which mimic backbone ß-turns and α-turns. Histidine residues are shown to serve as a mimic for Asn residues in side chain-backbone hydrogen bonded ring motifs. Illustrative examples from protein structures are considered.
Assuntos
Asparagina/química , Biologia Computacional , Glutamina/química , Domínios e Motivos de Interação entre Proteínas , Motivos de Aminoácidos , Histidina/química , Ligação de Hidrogênio , Estabilidade Proteica , Estrutura Secundária de Proteína , Proteínas/químicaRESUMO
Cross-strand disulfides bridge two cysteines in a registered pair of antiparallel ß-strands. A nonredundant data set comprising 5025 polypeptides containing 2311 disulfides was used to study cross-strand disulfides. Seventy-six cross-strand disulfides were found of which 75 and 1 occurred at non-hydrogen-bonded (NHB) and hydrogen-bonded (HB) registered pairs, respectively. Conformational analysis and modeling studies demonstrated that disulfide formation at HB pairs necessarily requires an extremely rare and positive χ¹ value for at least one of the cysteine residues. Disulfides at HB positions also have more unfavorable steric repulsion with the main chain. Thirteen pairs of disulfides were introduced in NHB and HB pairs in four model proteins: leucine binding protein (LBP), leucine, isoleucine, valine binding protein (LIVBP), maltose binding protein (MBP), and Top7. All mutants LIVBP T247C V331C showed disulfide formation either on purification, or on treatment with oxidants. Protein stability in both oxidized and reduced states of all mutants was measured. Relative to wild type, LBP and MBP mutants were destabilized with respect to chemical denaturation, although the sole exposed NHB LBP mutant showed an increase of 3.1°C in T(m). All Top7 mutants were characterized for stability through guanidinium thiocyanate chemical denaturation. Both exposed and two of the three buried NHB mutants were appreciably stabilized. All four HB Top7 mutants were destabilized (ΔΔG° = -3.3 to -6.7 kcal/mol). The data demonstrate that introduction of cross-strand disulfides at exposed NHB pairs is a robust method of improving protein stability. All four exposed Top7 disulfide mutants showed mild redox activity.
Assuntos
Peptídeos/química , Dicroísmo Circular , Cisteína/química , Dissulfetos/química , Ligação de Hidrogênio , Modelos Moleculares , Mutagênese Sítio-Dirigida , Oxirredução , Engenharia de Proteínas , Estabilidade Proteica , Estrutura Secundária de Proteína , Desdobramento de Proteína , Tiorredoxinas/químicaRESUMO
Aspartyl tRNA synthetase (AspRS), one of the 20 aminoacyl-tRNA synthetases, plays an important role in protein synthesis by catalyzing the aminoacylation reaction and synthesises Aspartyl-tRNA (tRNAAsp). A typical three-dimensional structure of AspRS comprises three distinct domains for the recognition of cognate tRNA and catalysis, namely, anti-codon binding domain/N-terminal domain, hinge domain and catalytic domain through their interactions with anti-codon loop, D-stem and acceptor arm of cognate tRNA, respectively. In this work, we have studied the structural characteristics of each domain of AspRS to understand the recognition mechanism of tRNAAsp using molecular dynamics simulations. The dynamics of AspRS-tRNAAsp complexes from E.coli (cognate and non-cognate), S.cerevisiae (cognate) and T.thermophilus (non-cognate) were compared to understand the differences in recognition of cognate and non-cognate tRNAs. Our results explain that the conformational changes associated with the recognition of tRNA occur only in the cognate complexes. Among the cognate complexes, the conformational changes in yeast AspRS are highly controlled during tRNAAsp recognition than that of in the E. coli AspRS. Moreover, the functional motions required for the tRNA recognition are observed only in the cognate complexes, and the conformational changes in AspRS and their recognition of tRNAAsp are organism specific.Communicated by Ramaswamy H. Sarma.
Assuntos
Aspartato-tRNA Ligase , Anticódon , Aspartato-tRNA Ligase/genética , Aspartato-tRNA Ligase/metabolismo , Sítios de Ligação , Escherichia coli/genética , Simulação de Dinâmica Molecular , RNA de Transferência de Ácido AspárticoRESUMO
To understand structural and thermodynamic features of disulfides within an alpha-helix, a non-redundant dataset comprising of 5025 polypeptide chains containing 2311 disulfides was examined. Thirty-five examples were found of intrahelical disulfides involving a CXXC motif between the N-Cap and third helical positions. GLY and PRO were the most common amino acids at positions 1 and 2, respectively. The N-Cap residue for disulfide bonded CXXC motifs had average (phi,psi) values of (-112 +/- 25.2 degrees , 106 +/- 25.4 degrees ). To further explore conformational requirements for intrahelical disulfides, CYS pairs were introduced at positions N-Cap-3; 1,4; 7,10 in two helices of an Escherichia coli thioredoxin mutant lacking its active site disulfide (nSS Trx). In both helices, disulfides formed spontaneously during purification only at positions N-Cap-3. Mutant stabilities were characterized by chemical denaturation studies (in both oxidized and reduced states) and differential scanning calorimetry (oxidized state only). All oxidized as well as reduced mutants were destabilized relative to nSS Trx. All mutants were redox active, but showed decreased activity relative to wild-type thioredoxin. Such engineered disulfides can be used to probe helix start sites in proteins of unknown structure and to introduce redox activity into proteins. Conversely, a protein with CYS residues at positions N-Cap and 3 of an alpha-helix is likely to have redox activity.
Assuntos
Motivos de Aminoácidos , Dissulfetos/química , Peptídeos/química , Estrutura Secundária de Proteína , Dicroísmo Circular , Cisteína/química , Cisteína/metabolismo , Bases de Dados de Proteínas , Insulina/química , Dados de Sequência Molecular , Mutagênese , Oxirredução , Peptídeos/genética , Desnaturação Proteica , Dobramento de Proteína , Termodinâmica , Tiorredoxinas/química , Tiorredoxinas/genéticaRESUMO
In the era of big data, the interplay of artificial and human intelligence is the demanding job to address the concerns involving exchange of decisions between both sides. Drug discovery is one of the key sources of the big data, which involves synergy among various computational methods to achieve a clinical success. Rightful acquisition, mining and analysis of the data related to ligand and targets are crucial to accomplish reliable outcomes in the entire process. Novel designing and screening tactics are necessary to substantiate a potent and efficient lead compounds. Such methods are emphasized and portrayed in the current review targeting protein-ligand and protein-protein interactions involved in various diseases with potential applications.
Assuntos
Antineoplásicos/química , Antivirais/química , Dengue/tratamento farmacológico , Desenho de Fármacos , Flavonoides/química , Neoplasias/tratamento farmacológico , Antineoplásicos/uso terapêutico , Antivirais/uso terapêutico , Biologia Computacional/métodos , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , RNA Polimerases Dirigidas por DNA/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Dengue/metabolismo , Dengue/virologia , Descoberta de Drogas/métodos , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Receptores ErbB/metabolismo , Flavonoides/uso terapêutico , Humanos , Ligantes , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Mapeamento de Interação de Proteínas , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
Inter-specific hybrids were produced between the threatened catfish species Mystus gulio x Mystus montanus. The differences in percentage of fertilization and hatching between control and interspecies were significant. The survival of hybrid was significantly lower (24.80 +/- 4.3%) when compared to control (95.1 +/- 3.5%). Time difference in yolk absorption by hybrid (73.30 h) was higher than that of control (72 h). When compared to interspecific fertilized egg the hatching time (24-25 h) and viability of larvae of the control were significantly better. In hybrids more deformed hatchlings (52.7 +/- 4.2 %) were noticed than the control (24.80 +/- 4.3%).
Assuntos
Peixes-Gato/embriologia , Embrião não Mamífero/embriologia , Fertilização in vitro/métodos , Hibridização Genética , Animais , Peixes-Gato/genética , Proliferação de Células , Cruzamentos Genéticos , Embrião não Mamífero/anormalidades , Embrião não Mamífero/citologia , Feminino , Água Doce , Masculino , Fatores de Tempo , Zigoto/citologia , Zigoto/crescimento & desenvolvimentoRESUMO
Abbreviations HA Hemagglutinin MD Molecular Dynamics MM-PBSA Molecular Mechanics Poisson-Boltzmann Surface Area NA Neuraminidase NAMD Nanoscale Molecular Dynamic Simulation PMEMD Particle Mesh Ewald Molecular Dynamics RMSD Root-Mean-Square Deviation RMSF Root-Mean-Square Fluctuation SIA sialic acid VMD Visual Molecular Dynamics Communicated by Ramaswamy H. Sarma.
Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Virus da Influenza A Subtipo H5N1/química , Ácido N-Acetilneuramínico/química , Sítios de Ligação , Ligação de Hidrogênio , Ligantes , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Ácido N-Acetilneuramínico/análogos & derivadosRESUMO
Disulfide bonds play an important role in protein stability and function. Here, we describe a general procedure for generating disulfide-linked dimers and multimers of proteins of known crystal structures. An algorithm was developed to predict sites in a protein compatible with intermolecular disulfide formation with neighboring molecules in the crystal lattice. A database analysis was carried out on 46 PDB coordinates to verify the general applicability of this algorithm to predict intermolecular disulfide linkages. On the basis of the predictions from this algorithm, mutants were constructed and characterized for a model protein, thioredoxin. Of the five mutants, as predicted, in solution four formed disulfide-linked dimers while one formed polymers. Thermal and chemical denaturation studies on these mutant thioredoxins showed that three of the four dimeric mutants had similar stability to wild-type thioredoxin while one had lower stability. Three of the mutant dimers crystallized readily (in four to seven days) in contrast to the wild-type protein, which is particularly difficult to crystallize and takes more than a month to form diffraction-quality crystals. In two of the three cases, the structure of the dimer was exactly as predicted by the algorithm, while in the third case the relative orientation of the monomers in the dimer was different from the predicted one. This methodology can be used to enhance protein crystallizability, modulate the oligomerization state and to produce linear chains or ordered three-dimensional protein arrays.
Assuntos
Algoritmos , Dissulfetos/química , Estrutura Quaternária de Proteína , Tiorredoxinas/química , Animais , Cristalografia por Raios X , Dimerização , Insulina/química , Insulina/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Oxirredução , Desnaturação Proteica , Termodinâmica , Tiorredoxinas/metabolismoRESUMO
When incorporated into a polypeptide chain, proline (Pro) differs from all other naturally occurring amino acid residues in two important respects. The phi dihedral angle of Pro is constrained to values close to -65 degrees and Pro lacks an amide hydrogen. Consequently, mutations which result in introduction of Pro can significantly affect protein stability. In the present work, we describe a procedure to accurately predict the effect of Pro introduction on protein thermodynamic stability. Seventy-seven of the 97 non-Pro amino acid residues in the model protein, CcdB, were individually mutated to Pro, and the in vivo activity of each mutant was characterized. A decision tree to classify the mutation as perturbing or nonperturbing was created by correlating stereochemical properties of mutants to activity data. The stereochemical properties including main chain dihedral angle phi and main chain amide H-bonds (hydrogen bonds) were determined from 3D models of the mutant proteins built using MODELLER. We assessed the performance of the decision tree on a large dataset of 163 single-site Pro mutations of T4 lysozyme, 74 nsSNPs, and 52 other Pro substitutions from the literature. The overall accuracy of this algorithm was found to be 81% in the case of CcdB, 77% in the case of lysozyme, 76% in the case of nsSNPs, and 71% in the case of other Pro substitution data. The accuracy of Pro scanning mutagenesis for secondary structure assignment was also assessed and found to be at best 69%. Our prediction procedure will be useful in annotating uncharacterized nsSNPs of disease-associated proteins and for protein engineering and design.
Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/ultraestrutura , Toxinas Bacterianas/química , Modelos Químicos , Modelos Moleculares , Mutagênese Sítio-Dirigida/métodos , Análise de Sequência de Proteína/métodos , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Simulação por Computador , Dados de Sequência Molecular , Mutação , Conformação Proteica , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
The amino acid Pro is more rigid than other naturally occurring amino acids and, in proteins, lacks an amide hydrogen. To understand the structural and thermodynamic effects of Pro substitutions, it was introduced at 13 different positions in four different proteins, leucine-isoleucine-valine binding protein, maltose binding protein, ribose binding protein, and thioredoxin. Three of the maltose binding protein mutants were characterized by X-ray crystallography to confirm that no structural changes had occurred upon mutation. In the remaining cases, fluorescence and CD spectroscopy were used to show the absence of structural change. Stabilities of wild type and mutant proteins were characterized by chemical denaturation at neutral pH and by differential scanning calorimetry as a function of pH. The mutants did not show enhanced stability with respect to chemical denaturation at room temperature. However, 6 of the 13 single mutants showed a small but significant increase in the free energy of thermal unfolding in the range of 0.3-2.4 kcal/mol, 2 mutants showed no change, and 5 were destabilized. In five of the six cases, the stabilization was because of reduced entropy of unfolding. However, the magnitude of the reduction in entropy of unfolding was typically several fold larger than the theoretical estimate of -4 cal K(-1) mol(-1) derived from the relative areas in the Ramachandran map accessible to Pro and Ala residues, respectively. Two double mutants were constructed. In both cases, the effects of the single mutations on the free energy of thermal unfolding were nonadditive.
Assuntos
Proteínas de Transporte/química , Proteínas de Escherichia coli/química , Proteínas Periplásmicas de Ligação/química , Prolina/química , Desnaturação Proteica , Tiorredoxinas/química , Substituição de Aminoácidos , Proteínas de Transporte/genética , Dicroísmo Circular , Cristalografia por Raios X , Entropia , Escherichia coli/química , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Modelos Moleculares , Mutagênese Sítio-Dirigida , Mutação , Proteínas Periplásmicas de Ligação/genética , Conformação Proteica , Desnaturação Proteica/efeitos dos fármacos , Dobramento de Proteína , Proteínas Recombinantes de Fusão/química , Espectrometria de Fluorescência , Relação Estrutura-Atividade , Temperatura , Termodinâmica , Tiorredoxinas/genéticaRESUMO
Motifs that are evolutionarily conserved in proteins are crucial to their structure and function. In one of our earlier studies, we demonstrated that the conserved motifs occurring invariantly across several organisms could act as structural determinants of the proteins. We observed the abundance of glycyl residues in these invariantly conserved motifs. The role of glycyl residues in highly conserved motifs has not been studied extensively. Thus, it would be interesting to examine the structural perturbations induced by mutation in these conserved glycyl sites. In this work, we selected a representative set of invariant signature (IS) peptides for which both the PDB structure and mutation information was available. We thoroughly analyzed the conformational features of the glycyl sites and their local interactions with the surrounding residues. Using Ramachandran angles, we showed that the glycyl residues occurring in these IS peptides, which have undergone mutation, occurred more often in the L-disallowed as compared with the L-allowed region of the Ramachandran plot. Short range contacts around the mutation site were analyzed to study the steric effects. With the results obtained from our analysis, we hypothesize that any change of activity arising because of such mutations must be attributed to the long-range interaction(s) of the new residue if the glycyl residue in the IS peptide occurred in the L-allowed region of the Ramachandran plot. However, the mutation of those conserved glycyl residues that occurred in the L-disallowed region of the Ramachandran plot might lead to an altered activity of the protein as a result of an altered conformation of the backbone in the immediate vicinity of the glycyl residue, in addition to long range effects arising from the long side chains of the new residue. Thus, the loss of activity because of mutation in the conserved glycyl site might either relate to long range interactions or to local perturbations around the site depending upon the conformational preference of the glycyl residue.
Assuntos
Motivos de Aminoácidos , Proteínas de Bactérias/química , Sequência Conservada , Glicina/química , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/genética , Sequência Conservada/genética , Escherichia coli , Glicina/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutação , Plasmodium falciparum , Células Procarióticas , Dobramento de Proteína , Sulfolobus , Vibrio choleraeRESUMO
Using a data set of 454 crystal structures of peptides and 80 crystal structures of non-homologous proteins solved at ultra high resolution of 1.2 A or better we have analyzed the occurrence of disallowed Ramachandran (phi, psi) angles. Out of 1492 and 13508 non-glycyl residues in peptides and proteins respectively 12 and 76 residues in the two datasets adopt clearly disallowed combinations of Ramachandran angles. These examples include a number of conformational points which are far away from any of the allowed regions in the Ramachandran map. According to the Ramachandran map a given (phi, psi) combination is considered disallowed when two non-bonded atoms in a system of two-linked peptide units with ideal geometry are prohibitively proximal in space. However, analysis of the disallowed conformations in peptide and protein structures reveals that none of the observations of disallowed conformations in the crystal structures correspond to a short contact between non-bonded atoms. A further analysis of deviations of bond lengths and angles, from the ideal peptide geometry, at the residue positions of disallowed conformations in the crystal structures suggest that individual bond lengths and angles are all within acceptable limits. Thus, it appears that the rare tolerance of disallowed conformations is possible by gentle and acceptable deviations in a number of bond lengths and angles, from ideal geometry, over a series of bonds resulting in a net gross effect of acceptable non-bonded inter-atomic distances.
Assuntos
Proteínas de Bactérias/química , Peptídeos/química , Sequência de Aminoácidos , Biologia Computacional , Genoma , Dados de Sequência Molecular , Mycobacterium/química , Conformação ProteicaRESUMO
All steroid receptors possess a bipartite nuclear localization signal sequence (NLS) that localizes within the second zinc finger of their DNA-binding domain. Fine-structure mapping of the rat glucocorticoid receptor (rGS) NLS identified a composite signal composed of three distinct proto-NLSs that function effectively when present in unique pairs. At least one of the rGR proto-NLSs appears to influence receptor trafficking within the nucleus, as revealed by a unique nuclear staining pattern of receptors possessing a point mutation (i.e., arginine at position 496; R496), at proto-NLS, pNLS-2. Specifically, carboxyl-terminal-truncated rGRs possessing various point mutations at R496 localized within a limited number of large foci in nuclei of transiently transfected COS-1 cells. R496 mutations did not affect subnuclear targeting when present in full-length rGR, reflecting a protective effect of the receptor's ligand-binding domain that can be exerted in cis and in trans. The effects of rGR R496 mutations on subnuclear targeting were not autonomous because we also observed a coincident localization of hsp70, the 70-kDa heat shock protein, within nuclear foci that include r496 mutant receptors. The elimination of R496 mistargeting by overexpression of an hsp70 partner (i.e., the DnaJ homologue, HDJ-2/HSDJ) suggests that the hsp70/DnaJ chaperone system is mobilized to specific sites within the nucleus in response to inappropriate targeting or folding of specific mutant receptors. HDJ-2/HSDJ overexpression also corrects defective transactivation and transrepression activity of R496 mutant GRs. Thus, molecular chaperones, such as members of the hsp70 and DnaJ families, may survey the nucleus for misfolded proteins and actively participate in their refolding into biologically active conformational states.
Assuntos
Proteínas de Transporte , Proteínas de Choque Térmico , Proteínas Nucleares/metabolismo , Receptores de Glucocorticoides/metabolismo , Ativação Transcricional , Dedos de Zinco , Sequência de Aminoácidos , Animais , Arginina , Sítios de Ligação , Transporte Biológico , Células COS , Núcleo Celular/metabolismo , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP40 , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Ligantes , Dados de Sequência Molecular , Mutação , Sinais de Localização Nuclear , Proteínas Nucleares/genética , Ratos , Receptores de Glucocorticoides/genética , TransfecçãoRESUMO
The functional significance of evolutionarily conserved motifs/patterns of short regions in proteins is well documented. Although a large number of sequences are conserved, only a small fraction of these are invariant across several organisms. Here, we have examined the structural features of the functionally important peptide sequences, which have been found invariant across diverse bacterial genera. Ramachandran angles (phi,psi) have been used to analyze the conformation, folding patterns and geometrical location (buried/exposed) of these invariant peptides in different crystal structures harboring these sequences. The analysis indicates that the peptides preferred a single conformation in different protein structures, with the exception of only a few longer peptides that exhibited some conformational variability. In addition, it is noticed that the variability of conformation occurs mainly due to flipping of peptide units about the virtual C(alpha)...C(alpha) bond. However, for a given invariant peptide, the folding patterns are found to be similar in almost all the cases. Over and above, such peptides are found to be buried in the protein core. Thus, we can safely conclude that these invariant peptides are structurally important for the proteins, since they acquire unique structures across different proteins and can act as structural determinants (SD) of the proteins. The location of these SD peptides on the protein chain indicated that most of them are clustered towards the N-terminal and middle region of the protein with the C-terminal region exhibiting low preference. Another feature that emerges out of this study is that some of these SD peptides can also play the roles of "fold boundaries" or "hinge nucleus" in the protein structure. The study indicates that these SD peptides may act as chain-reversal signatures, guiding the proteins to adopt appropriate folds. In some cases the invariant signature peptides may also act as folding nuclei (FN) of the proteins.
Assuntos
Proteínas de Bactérias/química , Sequência Conservada , Genoma Bacteriano , Dobramento de Proteína , Sequência de Aminoácidos , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Secundária de Proteína , Estrutura Terciária de ProteínaRESUMO
An analysis of the nature and distribution of disallowed Ramachandran conformations of amino acid residues observed in high resolution protein crystal structures has been carried out. A data set consisting of 110 high resolution, non-homologous, protein crystal structures from the Brookhaven Protein Data Bank was examined. The data set consisted of a total of 18,708 non-Gly residues, which were characterized on the basis of their backbone dihedral angles (phi, psi). Residues falling outside the defined "broad allowed limits" on the Ramachandran map were chosen and the reported B-factor value of the alpha-carbon atom was used to further select well defined disallowed conformations. The conformations of the selected 66 disallowed residues clustered in distinct regions of the Ramachandran map indicating that specific phi, psi angle distortions are preferred under compulsions imposed by local constraints. The distribution of various amino acid residues in the disallowed residue data set showed a predominance of small polar/charged residues, with bulky hydrophobic residues being infrequent. As a further check, for all the 66 cases non-hydrogen van der Waals short contacts in the protein structures were evaluated and compared with the ideal "Ala-dipeptide" constructed using disallowed dihedral angle (phi, psi) values. The analysis reveals that short contacts are eliminated in most cases by local distortions of bond angles. An analysis of the conformation of the identified disallowed residues in related protein structures reveals instances of conservation of unusual stereochemistry.
Assuntos
Aminoácidos/química , Conformação Proteica , Proteínas/química , Cristalografia por Raios X , Bases de Dados Factuais , Estrutura MolecularRESUMO
The occurrence of an eight-residue long segment of polypeptide chain in collagen helical conformation has been detected in bacteriochlorophyll a-protein by the application of an algorithm for identifying secondary structures in globular proteins from their alpha-carbon positions. This segment spans residues 277 to 284 of the protein and is the longest known stretch of collagen helix to be observed in globular proteins.
Assuntos
Proteínas de Bactérias , Colágeno , Sequência de Aminoácidos , Matemática , Modelos Moleculares , Conformação ProteicaRESUMO
A structure consisting of the polyproline-II or collagen-like helix immediately succeeded by a beta-turn is seen in several synthetic peptides and has been suggested to be the conformational requirement for proline hydroxylation in nascent procollagen. Using a simple algorithm for detecting secondary structures, we have analysed crystal structure data on 40 globular proteins and have found eight examples of the collagen-helix + beta-turn supersecondary structure in 15 proteins that contain the collagen-like helical segments.
Assuntos
Conformação Proteica , Algoritmos , Sequência de Aminoácidos , Colágeno , Dados de Sequência MolecularRESUMO
A super-secondary structural motif comprising two orthogonally oriented beta-strands connected by short linking segments of less than or equal to 5 residues has been identified from a data set of 65 independent protein crystal structures. Of the 42 examples from 14 proteins, a vast majority have only a single residue as the linking element. Analysis of the conformational angles at the junction reveals that the recently described type VIII beta-turn occurs frequently at the connecting hinge, while the type II beta-turn is also fairly common.