Calcium Ion-Induced Enzyme Dissociation and the Appearance of Enzyme Activity in Human Plasma Factor XIII

Human plasma Factor XIII (F. XIII) complex is composed of two types of noncovalently linked polypeptide chains termed a and b; only the a chain possesses catalytic potential. Platelet Factor XIII is comprised solely of a chains which are identical to those found in plasma. In this study platelets were utilized as a source of unbound a chains to characterize F. XIII (a chain)-binding activity in plasma and its subfractions. Upon exclusion chromatography of unheated plasma or of an unheated ammonium sulfate (20% sat.) subfraction, F. XIII activity emerged in a peak corresponding to a mol. wt. of < 500,000 (region 1). If these samples had first been heated at 60° to precipitate fibrinogen, F. XIII was eluted in a peak corresponding to a mol wt. of about 300,000 (region 2). Chromatography of the platelet zymogen alone resulted in a F. XIII peak corresponding in position to that of monomeric a chain (mol. wt. 80,000, region 3).Exclusion chromatography of the unheated ammonium sulfate fraction yielded, in addition to the F. XIII peak in region 1, a protein peak (peak II) in region 2 which contained no F. XIII. When peak II was mixed with platelet F. XIII, and again subjected to exclusion chromatography, the platelet F. XIII peak shifted from its expected position in region 3 and emerged instead in region 2 ; this behavior demonstrated F. XIII (a chain)-binding activity within peak II. The same chromatographic shift was observed in mixtures of platelet F. XIII and normal plasma or that from a patient with congenital F. XIII (a chain) deficiency. Immunochemical analyses of chromatographic fractions indicated that a chain-binding was due to complexing of a chains with freely circulating b chains. Since a chains and b chains have different biosynthetic sites we conclude that b chains serve as an extracellular F. XIII (a chain)-binding protein.Supported by NHLI grant HL-11409.

Download Full-text

Purification and Immunochemical Characterization of Human Plasma Factor XIII

Pathophysiology of Haemostasis and Thrombosis ◽

10.1159/000214153 ◽

1976 ◽

Vol 5 (6) ◽

pp. 329-340 ◽

Cited By ~ 2

Author(s):

M. Kazama ◽

Jan McDonagh ◽

R.H. Wagner ◽

R.D. Langdell ◽

R.P. McDonagh

Keyword(s):

Human Plasma ◽

Factor Xiii ◽

Immunochemical Characterization ◽

Plasma Factor

Download Full-text

PLASMA FACTOR XIII AND ACTIVATION PROCESS ANALYZED BY ELISA METHOD FOR A2B2 COMPLEX.

10.1055/s-0038-1643306 ◽

1987 ◽

Author(s):

K Koike ◽

M Hada ◽

H Yorifuji ◽

S Ikematsu ◽

K Fukutake ◽

...

Keyword(s):

Calcium Ion ◽

Factor Xiii ◽

Molecular Conformation ◽

Fibrin Clot ◽

Activation Process ◽

Measurement Sensitivity ◽

B Subunit ◽

Plasma Factor ◽

A Subunit ◽

Subunit B

Factor XIII induces the crosslinking of fibrin at terminal stage of blood coagulation. New ELISA methods of a-subunit, b-subunit and a2b2 complex of factor XIII were developed by the author and the following results concerning the activation process of factor XIII were obtained. New ELISA methods for a-subunit, b-subunit and a2b2 complex of factor XIII were specific with high sensitivities on each items and indicated the measurement capacity for simultaneous quantitation of many samples, more over we looked upon the dissociation of a2b2 complex with these methods to able to analyze at the subunit levels of factor XIII in details. When a-subunit dimer of platelets was discharged into the plasma by the use of freezing and thawing method on PRP, it was more easily affected to lose their antigeniety than that of a2b2 complex, a-subunit levels in the plasma of congenital factor XIII deficiencies were measured in very low concentration or below the measurement sensitivity of this ELISA, b-subunit levels in the same plasma were indicated around the half of normal levels. These results were as same as another immunological method. It was suggested that the molecular conformation of a-subunit could be changed by the addition of thrombin in high concentration and consequently a-subunit with thrombin was modifyed to show high antigeniety. It was observed that a2b2 complex was dissociated by the addition of thrombin without calcium ion, and the process of this dissociation of a2b2 complex was remarkably accelerated by the addition of calcium ion. Because a-subunit and b-subunit were adsorved on fibrin clot, it might be presumed that the crosslinking of fibrin molecules could be accelerated locally on the surface of fibrin clot.

Download Full-text

The Activation of Plasma Factor XIII with the Snake Venom Enzymes Ancrod and Batroxobin Marajoensis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651479 ◽

1977 ◽

Vol 38 (02) ◽

pp. 0438-0446 ◽

Cited By ~ 9

Author(s):

M Walter ◽

D Nyman ◽

V Krajnc ◽

F Duckert

Keyword(s):

Human Plasma ◽

Snake Venom ◽

Factor Xiii ◽

Test System ◽

Plasma Factor

SummaryThe snake venom enzymes Ancrod and Batroxobin marajoensis are able to activate human plasma factor XIII as shown by the formation of the γ-dimers. The concentration of γ-dimers increases with the concentration of the activating enzymes. Factor XIII activated by Ancrod or Batroxobin marajoensis is, however, unable to catalyse the incorporation of the amine dansylcadaverine into casein. The partially activated factor XIII is therefore not demonstrable by means of the artificial test system. This factor XIII loses little activity and remains activable by thrombin.

Download Full-text

A Pasteurised Concentrate of Human Plasma Factor XIII for Therapeutic Use

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661573 ◽

1986 ◽

Vol 55 (03) ◽

pp. 402-405 ◽

Cited By ~ 5

Author(s):

L Winkelman ◽

G E Sims ◽

M E Haddon ◽

D R Evans ◽

J K Smith

Keyword(s):

Human Plasma ◽

Intravenous Injection ◽

Plasma Proteins ◽

Factor Xiii ◽

Sodium Citrate ◽

Therapeutic Use ◽

Freeze Dried ◽

Plasma Factor ◽

Ethanol Fraction

SummaryA therapeutic concentrate of factor XIII containing both A and B sub-units has been prepared from 300 kg pools of human plasma. The process starts from a cold-ethanol fraction from cryoprecipitate supernatant and therefore does not interfere with the recovery of other clinically valuable plasma proteins. Factor XIII is purified approximately 600-fold from plasma by precipitation with sodium citrate and by the removal of fibrinogen by brief heating. The product has been pasteurised in sorbitol solution to inactivate blood-borne viruses, ultrafiltered to remove sorbitol, adsorbed with bentonite and freeze-dried in a formulation meeting requirements for intravenous injection.

Download Full-text

The Effect of Acetaldehyde on Human Plasma Factor XIII Function

Digestive Diseases and Sciences ◽

10.1007/s10620-007-9754-z ◽

2007 ◽

Vol 52 (12) ◽

pp. 3488-3492 ◽

Cited By ~ 2

Author(s):

Elizabeth A. Suchocki ◽

Arthur S. Brecher

Keyword(s):

Human Plasma ◽

Factor Xiii ◽

Plasma Factor

Download Full-text

The amino acid sequence around the reactive cysteine residue in human plasma Factor XIII (Short Communication)

Biochemical Journal ◽

10.1042/bj1350901 ◽

1973 ◽

Vol 135 (4) ◽

pp. 901-903 ◽

Cited By ~ 34

Author(s):

J. John Holbrook ◽

Rodney D. Cooke ◽

I. Barry Kingston

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Human Plasma ◽

Factor Xiii ◽

Short Communication ◽

Cysteine Residue ◽

Α Subunit ◽

Plasma Factor

1. Activated human plasma factor XIII was 65% inhibited by iodo[14C]acetate and incorporated 0.6 mol of label into the α subunit, which eventually was allowed to form a precipitate. 2. All the label was recovered as S-carboxymethylcysteine in a tetra-peptide of sequence Gly-Gln-Cys-Trp.

Download Full-text