Abstract
A new simple and efficient purification method for alpha 2-antiplasmin is described that is based on the interaction between alpha 2-antiplasmin and a fragment from elastase-digested plasminogen constituting the three N-terminal triple-loop structures in the plasmin A-chain (LBSI). After a single-step adsorption of the alpha 2-antiplasmin from plasminogen-depleted plasma to LBSI-Sepharose and elution with 6-aminohexanoic acid, an 80-90% pure preparation with a yield of 50-60% is obtained. The major impurity is fibrinogen, which can easily be removed by gel filtration, and, as a result, a homogeneous fully active alpha 2-antiplasmin preparation is obtained that has the same properties as previously described for alpha 2-antiplasmin. Evidence is put forward that a form of alpha 2-antiplasmin with less affinity for the lysine-binding sites in plasminogen may exist, even in unfractionated plasma.
MeSH Terms
Chromatography, Affinity/methods
Chromatography, Gel
Humans
Pancreatic Elastase
Plasminogen
Sepharose
alpha-2-Antiplasmin/isolation & purification
Chemicals
alpha-2-Antiplasmin
Plasminogen
Sepharose
Pancreatic Elastase
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Wiman B
References (16)
16 references, click to expand
-
Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides.
Nature. 1967 Jun 24;214(5095):1302-4
PMID: 6056841
-
Quantitative immunoelectrophoresis of human serum proteins.
Clin Sci. 1968 Oct;35(2):403-13
PMID: 5721242
-
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.
J Biol Chem. 1969 Aug 25;244(16):4406-12
PMID: 5806584
-
Plasminogen: purification from human plasma by affinity chromatography.
Science. 1970 Dec 4;170(3962):1095-6
PMID: 5475635
-
Activation of human plasminogen by an insoluble derivative of urokinase. Structural changes of plasminogen in the course of activation to plasmin and demonstration of a possible intermediate compound.
Eur J Biochem. 1973 Jul 2;36(1):25-31
PMID: 4270055
-
Isolation and characterization of alpha2-plasmin inhibitor from human plasma. A novel proteinase inhibitor which inhibits activator-induced clot lysis.
J Biol Chem. 1976 Oct 10;251(19):5956-65
PMID: 134998
-
Identification and some properties of a new fast-reacting plasmin inhibitor in human plasma.
Eur J Biochem. 1976 Oct 1;69(1):209-16
PMID: 136345
-
The primary inhibitor of plasmin in human plasma.
Biochem J. 1976 Dec 1;159(3):545-53
PMID: 137718
-
Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma.
Eur J Biochem. 1977 Aug 15;78(1):19-26
PMID: 21075
-
Fast-acting plasmin inhibitor in human plasma.
Blood. 1978 Apr;51(4):563-9
PMID: 147116
-
On the kinetics of the reaction between human antiplasmin and plasmin.
Eur J Biochem. 1978 Mar 15;84(2):573-8
PMID: 147769
-
On the kinetics of the reaction between human antiplasmin and a low-molecular-weight form of plasmin.
Eur J Biochem. 1978 Jun 1;87(1):143-6
PMID: 149657
-
Molecular mechanism of physiological fibrinolysis.
Nature. 1978 Apr 6;272(5653):549-50
PMID: 151233
-
Purification and reaction mechanisms of the primary inhibitor of plasmin from human plasma.
Biochem J. 1978 Nov 1;175(2):635-41
PMID: 154322
-
On the specific interaction between the lysine-binding sites in plasmin and complementary sites in alpha2-antiplasmin and in fibrinogen.
Biochim Biophys Acta. 1979 Jul 25;579(1):142-54
PMID: 157166
-
On the mechanism of the reaction between human alpha 2-antiplasmin and plasmin.
J Biol Chem. 1979 Sep 25;254(18):9291-7
PMID: 158022