Conversion of Pig Prothrombin into Thrombin: Amino Acid Composition of the Active Enzyme

1974 ◽  
Vol 52 (4) ◽  
pp. 336-344 ◽  
Author(s):  
Antonio Rossi ◽  
José Roberto Giglio
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Column Chromatography ◽  
Active Enzyme ◽  
Terminal Residue

Preparation and activation of partially purified pig prothrombin are described. The enzyme, after column chromatography in Amberlite IRC-50, showed alanine as the N-terminal residue and an [Formula: see text] of 1.50 at pH 7.0 and 280 nm.The amino acid composition of pig thrombin is reported and compared with that of bovine origin. Pig thrombin showed 1.0 mol of isoleucine to 0.6 mol of threonine as N-terminals.The effects of storage of pig prothrombin are discussed.

scholarly journals Action of human lysosomal elastase on the oxidized B chain of insulin

1977 ◽  
Vol 161 (1) ◽  
pp. 13-16 ◽  
Author(s):  
A M J Blow
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Human Neutrophil ◽  
British Library ◽  
Carboxyl Group ◽  
Pancreatic Elastase ◽  
Terminal Residue

The specificity of action of the lysosomal elastase of human neutrophil leucocytes on the oxidized B chain of insulin is similar to that of pig pancreatic elastase, but is more directed towards valine than alanine as the residue contributing the carboxyl group of the cleaved bond. The most susceptible bonds are Val-12-Glu-13 and Val-18-Cys(O3H)-19. Other bonds hydrolysed are Ala-14-Leu-15, Ser-9-His-10 and Cys, (O3H3)-7-Gly-8. Tables listing amino acid composition, N-terminal residue, and yields of isolated peptides have been deposited as Supplementary Publication SUP 50 075 (8 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1977) 161, 1.

scholarly journals AMINO ACID COMPOSITION AND ELECTROPHORETIC PROPERTIES OF HYPERTENSIN I

1955 ◽  
Vol 102 (4) ◽  
pp. 435-440 ◽  
Author(s):  
Leonard T. Skeggs ◽  
Walton H. Marsh ◽  
Joseph R. Kahn ◽  
Norman P. Shumway
Keyword(s):  
Amino Acids ◽  
Quantitative Analysis ◽  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Isoelectric Point ◽  
Column Chromatography ◽  
Isoleucine Leucine ◽  
Ion Exchange Column

A preparation of hypertensin I was purified by countercurrent distribution and was shown to migrate as a single component in starch blocks at pH 9.3 and 4.2. It had an isoelectric point of 7.7. Quantitative analysis by ion exchange column chromatography showed eight amino acids in approximately unimolar proportion: aspartic, proline, valine, isoleucine, leucine, tyrosine, phenylalanine, and arginine. There were in addition two moles of histidine.

scholarly journals Purification and biochemical characterization of three major acidic proteins (BSP-A1, BSP-A2 and BSP-A3) from bovine seminal plasma

1987 ◽  
Vol 241 (3) ◽  
pp. 685-692 ◽  
Author(s):  
P Manjunath ◽  
M R Sairam
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Seminal Plasma ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Terminal Residue ◽  
Acidic Proteins

Three major acidic proteins of bovine seminal plasma, BSP-A1, BSP-A2 and BSP-A3, were purified to homogeneity, by employing fast protein liquid chromatography, gel filtration and h.p.l.c. The proteins were purified on the basis of their stimulatory effect on the basal release of gonadotropins by rat anterior-pituitary cells in culture. All three proteins migrated as distinct single bands in the presence or absence of 2-mercaptoethanol in SDS/polyacrylamide-gel electrophoresis. Their Mr values were estimated to be between 15,000 and 16,500 by SDS/polyacrylamide-gel electrophoresis. Similar Mr estimates were obtained when they were subjected to gel filtration on a calibrated column of Sephadex G-75 equilibrated in 0.05 M-acetic acid, pH 3.0. However, BSP-A1 and BSP-A2 were eluted as aggregated molecules (Mr 60,000-120,000) during gel filtration on Sephadex G-200 equilibrated in 0.05 M-NH4HCO3, pH 8.5, or phosphate buffer, pH 7.0, containing 0.15 M-NaCl. In the presence of 8 M-urea both BSP-A1 and BSP-A2 were eluted at positions corresponding to Mr values of 17,000-20,000. BSP-A1 and BSP-A2 had an identical amino acid composition, which differed largely from that of BSP-A3. All three proteins contained aspartic acid as the N-terminal residue, and cysteine was identified as the C-terminal residue. BSP-A1 and BSP-A2 are glycoproteins containing galactosamine, sialic acid and neutral sugars, but BSP-A3 did not contain any covalently attached sugars. Whereas BSP-A2 and BSP-A3 were eluted unadsorbed, BSP-A1 bound to wheat-germ lectin-Sepharose 6MB and could be eluted by the competing sugar N-acetyl-D-glucosamine. Treatment of BSP-A1 and BSP-A2 with trypsin resulted in complete loss of gonadotropin-release activity, but BSP-A3 retained full activity. Antibody raised against BSP-A1 did not cross-react with BSP-A3, or vice versa. All these properties indicated marked structural differences between BSP-A3 and BSP-A1 (or BSP-A2). On the basis of amino acid composition it was concluded that BSP-A1, BSP-A2 and BSP-A3 are the same as the gonadostatins [Esch, Ling, Bohlen, Ying & Guillemin (1983) Biochem. Biophys. Res. Commun. 113, 861-867].

scholarly journals Action of human cathepsin G on the oxidized B chain of insulin

1977 ◽  
Vol 161 (1) ◽  
pp. 17-19 ◽  
Author(s):  
A M J Blow ◽  
A J Barrett
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Human Neutrophil ◽  
Cathepsin G ◽  
British Library ◽  
Neutral Proteinase ◽  
Terminal Residue ◽  
Human Cathepsin ◽  
Chymotrypsin A

The specificity of cathepsin G, a serine neutral proteinase from human neutrophil leucotyes, was determine dby its action on the insulin B chain. The most susceptible bonds were Phe-24-Phe-25, Leu-15-Tyr-16 and Tyr-16-Leu-17. Other bonds hydrolysed were Leu-6-Cys(O3H)-7, Leu-11-Val-12, Leu-17-Val-18 and Phe-25-Tyr-26. These results suggest that the specificity of cathespin G is closer to that of pig chymotrypsin C than ox Chymotrypsin A. Tables listing amino acid composition, N-terminal residue, and yields of isolated peptides have been deposited as Supplementary Publication SUP 50 075 (8 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7B2, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1977) 161,1.

Human pituitary thyrotropin. Isolation and characterization of the hormone and its subunits

1977 ◽  
Vol 55 (7) ◽  
pp. 747-754 ◽  
Author(s):  
M. R. Sairam ◽  
Choh Hao Li
Keyword(s):  
Biological Activity ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Disc Electrophoresis ◽  
Carbohydrate Content ◽  
Human Pituitary ◽  
Terminal Residue ◽  
Isolation And Characterization

Procedures have been described for the isolation of highly purified thyrotropin from frozen or acetone-preserved glands or from side fractions of somatotropin isolation and for the separation of its α and β subunits. The products have been characterized by terminal residue analyses, amino acid composition, carbohydrate content, disc electrophoresis, ultracentrifugation, and biological activity.

Hydroxyproline-rich protein in the capsule of a strain of Staphylococcus aureus

1981 ◽  
Vol 27 (9) ◽  
pp. 955-958 ◽  
Author(s):  
Yukio Usui ◽  
Kosaku Yoshida ◽  
C. L. San Clemente
Keyword(s):  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Sodium Chloride ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Column Chromatography ◽  
Protein Fraction ◽  
Oxygen Supply ◽  
Total Amino Acid

A protein fraction containing a high amount of hydroxyproline was found in the capsule of the Smith diffuse strain (encapsulated) when compared with that of dialysate medium. This content varied depending upon temperature, oxygen supply, sodium chloride, and carbohydrates. The hyroxyproline content of the single proteinaceous fraction, obtained by Sephadex G-200 column chromatography, was one fourth of the total amino acid composition.

THE EFFECTS OF D-AMINO ACIDS ON ALCALIGENES FECALIS

1959 ◽  
Vol 5 (4) ◽  
pp. 369-379 ◽  
Author(s):  
Cynthia Lark ◽  
K. G. Lark
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Column Chromatography ◽  
Cell Walls ◽  
Insoluble Fraction ◽  
Normal Cells ◽  
Basic Peptide ◽  
Peptide Unit

D-Amino acids have been found to induce protoplast-like forms (crescents) in Alcaligenes fecalis strain LB. This induction is similar in most respects to that by penicillin with the exception of the effects produced in tryptone. D-Amino acids appear to act synergistically with penicillin. The amino acid composition of cell walls of D-amino acid and penicillin-induced crescents as well as of normal cells has been examined by column chromatography. The main difference between normal and crescent cell walls consists of a reduction in the amount of the basic peptide unit of the phenol-insoluble fraction. These results are discussed in relation to the induction of protoplast-like forms.

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