The Amino Acid Sequence of Streptomyces griseus Trypsin. I. The Peptic Peptides

1973 ◽  
Vol 51 (7) ◽  
pp. 1077-1088 ◽  
Author(s):  
L. Jurášek ◽  
L. B. Smillie
Keyword(s):  
Amino Acid ◽  
Serine Proteases ◽  
System Analysis ◽  
Streptomyces Griseus ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Terminal Sequence ◽  
Unique Amino Acid ◽  
High Voltage Electrophoresis

Streptomyces griseus trypsin (S.G.T.) isolated from pronase was digested with pepsin. The peptic peptides were isolated by high-voltage electrophoresis on paper and ion-exchange chromatography on Chromobead P resin using the Technicon autoanalyzer system. Analysis of the purified peptides provides 28 unique amino acid sequences accounting for approximately 95% of the S.G.T. molecule. A portion of the residues not accounted for can be ascribed to free leucine, phenylalanine, tyrosine, and tryptophan present in the peptic digest. The NH2-terminal sequence of S.G.T. was shown to be Val–Val–Gly–Gly–Thr–Arg–Ala–Ala–Gln–Gly–Glu–Phe and is highly homologous with NH2-terminal sequences of other Asp–Ser–Gly serine proteases.

Serine protease isoforms of Deinagkistrodon acutus venom: cloning, sequencing and phylogenetic analysis

2001 ◽  
Vol 354 (1) ◽  
pp. 161-168 ◽  
Author(s):  
Ying-Ming WANG ◽  
Suei-Rong WANG ◽  
Inn-Ho TSAI
Keyword(s):  
Amino Acid ◽  
Serine Proteases ◽  
Parallel Evolution ◽  
Gel Filtration ◽  
Venom Gland ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Cdna Clones ◽  
Single Chain

The major coagulating fibrinogenase of Deinagkistrdon acutus venom, designated acutobin, was purified by anion-exchange chromatography, gel filtration and reverse-phase HPLC. Approximately 80% of its protein sequence was determined by sequencing the various fragments derived from CNBr cleavage and digestion with endoprotease. Extensive screening of the venom gland cDNA species after amplification by PCR resulted in the isolation of four distinct cDNA clones encoding acutobin and three other serine proteases, designated Dav-PA, Dav-KN and Dav-X. The complete amino acid sequences of these enzymes were deduced from the cDNA sequences. The amino-acid sequence of acutobin contains a single chain of 236 residues including four potential N-glycosylation sites. The purified acutobin (40kDa) contains approx. 30% carbohydrate by weight, which could be partly removed by N-glycanase. The phylogenetic tree of the complete amino acid sequences of 40 serine proteases from 18 species of Crotalinae shows functional clusters reflecting parallel evolution of the three major venom enzyme subtypes: coagulating enzymes, kininogenases and plasminogen activators. The possible structural elements responsible for the functional specificity of each subtype are discussed.

scholarly journals Studies on Marsupial Protein. II. Amino Acid Sequence of the -Chain of Haemoglobin from the Grey Kangaroo, Macropus Giganteus

1969 ◽  
Vol 22 (6) ◽  
pp. 1437 ◽  
Author(s):  
GM Air ◽  
EOP Thompson
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Sequence ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Tryptic Digestion ◽  
Grey Kangaroo

The amino acid sequence of the jS-chain of haemoglobin from M. giganteus has been determined. The soluble peptides formed by tryptic digestion were isolated by gel filtration, ion-exchange chromatography, and paper ionophoresis, and amino acid sequences determined by the "dansyl"-Edman procedure. Special procedures were necessary for three peptides which were insoluble.

Thermolytic digest of chicken pepsin

1981 ◽  
Vol 46 (8) ◽  
pp. 1994-2004 ◽  
Author(s):  
Miroslav Baudyš ◽  
Vladimír Kostka ◽  
Helena Keilová
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Linear Structure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence ◽  
High Voltage Electrophoresis ◽  
Final Purification

Chicken pepsin prepared by the activation of pepsinogen was digested with thermolysin. The thermolytic digest was fractionated by chromatography on Sephadex G-25 fine. Certain fractions were subsequently subjected to ion exchange chromatography on Dowex 50-X2. The final purification was effected by paper chromatography and high voltage electrophoresis. By these procedures a series of homogeneous peptides was obtained; of the latter 54 nonoverlapping (save for a few exceptions) peptides are described in this paper. These peptides in addition to the thermolytic peptides reported before represent 80% of the linear structure of the whole molecule. The N-terminal amino acid sequence of chicken pepsin is discussed from the viewpoint of the recent data obtained by the analysis of the thermolytic digest.

Carnivora: The Amino Acid Sequence of the Adult European Mink (Mustela lutreola, Mustelidae) Hemoglobins

1990 ◽  
Vol 45 (3-4) ◽  
pp. 223-228 ◽  
Author(s):  
Aftab Ahmed ◽  
Meeno Jahan ◽  
Gerhard Braumtzer
Keyword(s):  
Amino Acid ◽  
Gas Phase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Mustela Lutreola ◽  
European Mink ◽  
Globin Chains ◽  
The Family ◽  
Insight Into

Abstract The complete amino acid sequences of the hemoglobins from the adult European mink (Mustela lutreola) are presented. The erythrocytes contain two hemoglobin components and three globin chains. The isolation of globin chains achieved by ion-exchange chromatography on a column of CM -cellulose in 8 M urea buffer. The primary structure of globin chains and of the tryptic peptides determined in liquid-and gas-phase sequenators. The alignment of the a-and β-chains with those of reported sequences from other carnivora species belonging to the family Mustelidae may give an insight into the evolution of this molecule.

scholarly journals Two globin strains in the giant annelid extracellular haemoglobins

1987 ◽  
Vol 241 (2) ◽  
pp. 441-445 ◽  
Author(s):  
T Gotoh ◽  
F Shishikura ◽  
J W Snow ◽  
K I Ereifej ◽  
S N Vinogradov ◽  
...  
Keyword(s):  
Amino Acid ◽  
Lumbricus Terrestris ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Reverse Phase ◽  
Terminal Amino ◽  
Polypeptide Chains

The constituent polypeptide chains I, II, III and IV of the giant extracellular haemoglobin of the oligochaete Lumbricus terrestris were isolated by mono Q ion-exchange chromatography and C8 reverse-phase chromatography. The N-terminal amino acid sequences of Lumbricus chains I, III and IV were determined and aligned with those of Lumbricus chain II and the four chains of the extracellular haemoglobin of the polychaete Tylorrhynchus heterochaetus. Three invariant amino acid residues, Cys-7, Val-15 and Trp-19, were found to occur in the N-terminal segments (17-22 residues) of the eight chains of Lumbricus and Tylorrhynchus haemoglobins. In addition, it was found that the eight sequences could be separated into two groups: ‘A’, consisting of Lumbricus chains I and II and Tylorrhynchus chains I and IIA, having invariant Lys-14 and Lys-16, and ‘B’, consisting of Lumbricus chains III and IV and Tylorrhynchus IIB and IIC, having invariant Cys-6, Ser-8 and Asp-11. This result suggests that there are two strains of globin chain in the annelid extracellular haemoglobins.

The Amino Acid Sequence of Streptomyces griseus Trypsin. II The Tryptic Peptides

1974 ◽  
Vol 52 (5) ◽  
pp. 382-392 ◽  
Author(s):  
L. Jurášek ◽  
L. B. Smillie
Keyword(s):  
Soluble Fraction ◽  
Streptomyces Griseus ◽  
Paper Electrophoresis ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Insoluble Residue ◽  
Limited Extent ◽  
Terminal Sequence ◽  
Tryptic Peptides ◽  
Final Purification

Streptomyces griseus trypsin (S.G.T.) isolated from pronase was reduced, aminoethylated, and digested with trypsin. The soluble peptides were recovered and the insoluble residue redigested with chymotrypsin. Following recovery of the soluble fraction, the insoluble portion was in turn digested with α-lytic protease of Myxobacter 495. The three groups of soluble peptides were separately subjected to ion-exchange chromatography on the Technicon peptide analyzer and to final purification by high-voltage paper electrophoresis. Sequence analysis by the dansyl-Edman procedure provided unique sequences from the soluble tryptic peptides accounting for 65% of the S.G.T. molecule. Peptides obtained from the redigests of the insoluble residues accounted for an additional 20%.Tryptic digestion of dansylated S.G.T. yielded a unique α-NH2 dansylated peptide whose composition showed it to be the same as the NH2-terminal sequence previously postulated for this enzyme. The tryptic peptides isolated in this work have provided overlaps for many of the previously sequenced peptic peptides. Three continuous sequences of 49, 36, and 28 residues have been elucidated.Evidence has also been obtained that the NH2-terminal valine residue was, to a limited extent, aminoethylated during reaction of the reduced protein with ethylenimine.

scholarly journals Yeast phosphoglycerate mutate. Cyanogen bromide cleavage and amino acid sequence of an active-site peptide

1976 ◽  
Vol 153 (2) ◽  
pp. 145-149 ◽  
Author(s):  
L A Fothergill ◽  
G I Hodgson
Keyword(s):  
Amino Acid ◽  
Active Site ◽  
Cyanogen Bromide ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Phosphoglycerate Mutase ◽  
Large Fragment ◽  
X Ray ◽  
Cnbr Cleavage

The molecular weight and amino acid composition of phosphoglycerate mutase from yeast were determined. CNBr cleavage produced a large (190-residue) fragment and a small (60-residue) fragment. Tryptic and chymotryptic peptides derived from the large fragment were fractionated by ion-exchange chromatography. Peptides from two histidine-containing regions were isolated and the amino acid sequences were determined. Correlation of these data with X-ray-crystallographic evidence shows that the histidine residue in the sequence Arg-Leu Asn-Glu-Arg-His-Tyr-Gly-Asp-Leu-Glu-Gly-Lys is located at the active site.

The Amino Acid Sequence of Streptomyces griseus Protease A: The Peptic Peptides

1971 ◽  
Vol 49 (10) ◽  
pp. 1083-1097 ◽  
Author(s):  
P. Johnson ◽  
L. B. Smillie
Keyword(s):  
Amino Acid ◽  
Streptomyces Griseus ◽  
Disulfide Bridge ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Homologous Proteins ◽  
Acidic Peptide ◽  
Dowex 1 ◽  
Peptide Fractions

The peptic peptides of Streptomyces griseus Protease A (excluding the previously characterized disulfide bridge peptic peptides) were fractionated into basic, neutral, and neutral plus acidic peptide fractions by chromatography on Dowex 1 × 2. These three peptide fractions were then fractionated by cation-exchange chromatography on Chromobead P resin using the Technicon autoanalyzer system. Following further purifications on paper, the amino acid compositions and sequences of the peptic peptides were determined. The N-terminal sequence of Protease A has been identified as Ile–Ala–Gly–Gly–Glu–Ala. The numbers of amino acid residues obtained from the amino acid sequences reported are in agreement with those numbers obtained from amino acid analysis of the total protein in the cases of tryptophan, methionine, histidine, proline, phenylalanine, and glutamic acid. Some of the results suggest either the presence of nonidentical but highly homologous proteins in the Protease A preparation or the possibility of repeating sequences in a single molecular species.

scholarly journals Antigenicity of proteinases from Streptomyces griseus (pronase)

1970 ◽  
Vol 119 (2) ◽  
pp. 339-342 ◽  
Author(s):  
M. Trop ◽  
R. R. Avtalion ◽  
Z. Malik ◽  
A. Pinsky
Keyword(s):  
Amino Acid ◽  
Active Sites ◽  
Streptomyces Griseus ◽  
Ion Exchange Chromatography ◽  
Enzymic Activity ◽  
Exchange Chromatography ◽  
Soya Bean ◽  
Catalytic Site ◽  
Antigenic Sites ◽  
Bovine Trypsin

The five pronase fractions, A1, A2, B, C (trypsin-like), and D (elastolytic), obtained by ion-exchange chromatography, were found to be antigenically distinct. Antibodies to pronase inhibited the enzymic activity of each of the enzyme fractions. Pronase trypsin and bovine trypsin, although resembling each other in enzymic activity and in amino acid sequence around their active sites, did not cross-react antigenically with, nor was their enzymic activity inhibited by, the respective homologous antibodies. Inactivation of pronase trypsin by complexing with soya-bean inhibitor AA, was not associated with a decrease in capacity to precipitate with its antibody. It is assumed that the antigenic sites are located far enough from the catalytic site of the enzyme to allow it to precipitate immunologically even when the catalytic site was blocked.

Bovine Factors X1And X2: Activation Peptide Based Chromatographic Differences

1979 ◽  
Author(s):  
Takashi Morita ◽  
Craig Jackson
Keyword(s):  
Amino Acid ◽  
Anion Exchange ◽  
Gel Filtration ◽  
Heart Association ◽  
Personal Communication ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Factor X ◽  
Activation Peptide ◽  
Activation Peptides

Bovine Factor X is eluted in two forms (X1and X2) from anion exchange chromatographic columns. These two forms have indistinguishable amino acid compositions, molecular weights and specific activities. The amino acid sequences containing the γ-carboxyglutamic acid residues have been shown to be identical in X1 and X2(H. Morris, personal communication). An activation peptide is released from the N-terminal region of the heavy chain of Factor X by an activator from Russell’s viper venom. This peptide can be isolated after activation by gel filtration on Sephadex G-100 under nondenaturing conditions. The activation peptides from a mixture of Factors X1 and X2 were separated into two forms by anion-exchange chromatography. The activation peptide (AP1) which eluted first was shown to be derived from Factor X1. while the activation peptiae (AP2) which eluted second was shown to be derived from X2 on the basis of chromatographic separations carried out on Factors X1 and X2 separately. Factor Xa was eluted as a symmetrical single peak. On the basis of these and other data characterizing these products, we conclude that the difference between X1 and X2 are properties of the structures of the activation peptides. (Supported by a grant HL 12820 from the National Heart, Lung and Blood Institute. C.M.J. is an Established Investigator of the American Heart Association).

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