RESUMO
The complete primary structures of two hemoglobin components of the European Souslik (Citellus citellus) are presented. The two hemoglobins have identical alpha-chains but differ in the amino-acid sequence of their beta-chains. The chain separation was achieved by chromatography on carboxymethyl-cellulose CM-52. Amino-acid sequences were established by automatic liquid-phase and gas-phase Edman degradation of the globin chains, of their tryptic peptides and of a peptide resulting from acidic hydrolysis of an Asp-Pro bond in the alpha-chain. The differences between the two beta-chains are manifested in three amino-acid exchanges. The sequences are compared with those of human and European Marmot hemoglobins. Only few differences were found among hemoglobins of C. citellus and other representatives of Sciuromorpha.
Assuntos
Hemoglobinas/análise , Sciuridae/sangue , Sequência de Aminoácidos , Aminoácidos/análise , Animais , Globulinas/análise , Hidrólise , TripsinaRESUMO
The primary structure of the beta A- and beta B-chains from both main components of the tetrameric hemoglobins from carp is given. The sequences were found by automatic analysis in the sequenator using a hydrophilic and hydrophobic program with hydrophilic isothiocyanate. The beta-chains A and B differ in positions beta 13, beta 41, beta 122 and beta 143. In comparison to human beta-chains carp beta A- and beta B-chains show 66 and 67 residues, respectively (45%). Furthermore, we found one insertion in the GH-region, therefore the chains contain 147 amino acids. The sequence is discussed and in particular, the chains of tetrameric hemoglobins are compared to those of mammalia: beta A and beta B contain 3 and 4 cysteines, respectively, but the external cysteine beta 93 of mammalia is replaced by serine. In ;the N-terminal region in E-helix and G-helix an accumulation of mutations is found. Furthermore, we think the high oxidation of the hemoglobins from carp comes from an external cysteine in beta 31 and beta 143, respectively. Several aspects of an ATP-hemoglobin exchange and its function are discussed.
Assuntos
Aminoácidos/análise , Carpas/sangue , Cyprinidae/sangue , Hemoglobinas/análise , Sequência de Aminoácidos , Animais , Autoanálise , Fenômenos Químicos , Química , Cromatografia/métodos , Substâncias Macromoleculares , Fragmentos de Peptídeos/análiseRESUMO
In a previous publication, a new phospholipid was isolated from the keratinoidal layer of chicken gizzard, and on the basis of chemical, spectrometric (IR, NMR, UV) tests and mass analysis, the probable structure of the isolated phospholipid was proposed as 1,2-O-docosylidene-sn-glycero-3-phospho(1'-ribosyl)-ethanolamine (IX). In this paper, on the basis of probable structure, a new phospholipid was synthetized.
Assuntos
Moela das Aves/metabolismo , Fosfolipídeos/biossíntese , Animais , Galinhas , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Fosfolipídeos/síntese química , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
The primary structures of the beta A - and beta B-chains from the both main components of carp hemoglobins is given. The sequence was established by splitting with known methods and sequencing the products using the hydrophilic and hydrophobic programmes in the sequenator. The carp beta-chains differ from human beta-chains in the exchange of 66 and 67 amino acid residues, respectively; the beta A - beta B-chains differ by 4 amino acids (pos. no. 13, 41, 122 and 143). Both chains have one insertion in the position 121 or 122. The results are discussed.
Assuntos
Hemoglobinas , Sequência de Aminoácidos , Animais , Carpas , Substâncias Macromoleculares , Fragmentos de Peptídeos/análiseRESUMO
We have established several optimal conditions for qualitative and quantitative allantoin determination by applying Ehrlich's reagent. The limit of detection for allantoin determination amounts to 5 x 10(-6) mM. Allantoin is determined quantitatively by measuring the absorbance at 440 nm (from 300 to 1000 micrograms/ml). The color of the complex becomes stable by standing for 10 min at room temperature. We have used these conditions for allantoin determination in Agrostemma githago seed.
Assuntos
Alantoína/análise , Plantas/química , Benzaldeídos , Indicadores e Reagentes , Sementes/química , EspectrofotometriaRESUMO
A total of 18 60-day-old male Wistar rats were divided into three groups of six animals each. One group was fed a basal diet containing high levels of protein, fat, carbohydrate, vitamins, and minerals and separately a solution of 25% sucrose-32% ethyl alcohol (wt/vol). A second group was offered water as the only drinking fluid and a similar solid diet, except that carbohydrate replaced ethanol isocalorically. A third group was maintained on the basal diet ad libitum. All groups of animals were killed in a sober state after 6 months of chronic ethanol treatment and lipid analyses were performed on brain homogenates. Chronic treatment of the animals with ethanol produces statistically significant modification of the phospholipid and ganglioside patterns in rat brain. A statistically significant decrease of the total phospholipid content and of some of the investigated fractions, i.e., phosphatidylcholine and phosphatidylserine, as well as an increase of phosphatidylinositol were observed. Chronic alcohol consumption was associated with a statistically significant increase in the total amount of ganglioside in rat brain. An increase in most of the investigated ganglioside fractions was indicated but the difference was statistically significant only for trisialoganglioside GT1b. The amount of disialoganglioside GD1a in these brains was decreased after chronic intake of ethanol.
Assuntos
Encéfalo/metabolismo , Etanol/farmacologia , Gangliosídeos/metabolismo , Fosfolipídeos/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Cromatografia em Camada Fina , Cinética , Masculino , Ratos , Ratos EndogâmicosRESUMO
A new phospholipid (Mr = 653; C32H64NO10P; [alpha]22D: - 24) was isolated from the keratinoidal layer of chicken gizzard. After acid hydrolysis the following products were identified: a C22-saturated aldehyde, sn-glycerol 3-phosphate, ethanolamine and D(-)-ribose. The substance is stable to alkali, it has saturated character, and after hydrolysis with CH3OH/HCI, yields only C22-dimethyl acetal. It contains a free NH2 group, and methylation produces 2,3,4-tri-O-methyl-D-ribose. The probable structure of the isolated phospholipid is proposed as 1,2-O-do-cosylidene-sn-glycero-3-phospho-(1 inch-ribosyl)-ethanolamine.