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B Wiman

Publications and source records attributed to B Wiman.

At least 109 records · Page 6Linked to original sources
PubMed1982

Purification and characterization of human C1-esterase inhibitor.

A new purification method for C1-esterase inhibitor is described, which is essentially a three-step procedure: precipitation with poly(ethylene glycol), chromatography on DEAE-cellulose and hydrophobic interaction chromatography on hexyl-Sepharose. The final product is a single-chain glycoprotein with a molecular weight of about 100 000 and NH2-terminal asparagine. The molecule is fully active as judged by complex formation with C1s. Two of its three disulphide bridges can be easily reduced and S-carboxymethylated under non-denaturing conditions without loss of activity. However, at high dithioerythritol concentration the third disulphide bridge is also cleaved and accompanied by loss of the activity, indicating that this disulphide bridge is involved in maintaining the conformation around the reactive site in the inhibitor.

Amino Acids↗
PubMed1982

Purification and partial primary structure of cyanogen bromide fragments from human alpha 2-antiplasmin.

Human alpha 2-antiplasmin was digested with CNBr and the partial NH2-terminal amino acid sequences of nine purified fragments were determined. One of these sequences (12 residues) revealed homologies with that of residues 158 to 168 and with that of residues 316-327 in the antithrombin III sequence and with the corresponding sequences in alpha 1-antitrypsin and ovalbumin. One CNBr-fragment of 69 residues with blocked NH2-terminus was subdigested with trypsin and chymotrypsin and most of its sequence aligned. This peptide probably represents the major part of the previously reported COOH-terminal fragment of alpha 2-antiplasmin with Mr 8,000 (1). In this sequence no clear homology with antithrombin III, alpha 1-antitrypsin or ovalbumin could be detected.

Amino Acid Sequence↗
PubMed1980

Affinity-chromatographic purification of human alpha 2-antiplasmin.

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.

Chromatography, Affinity↗
PubMed1979

On the specific interaction between the lysine-binding sites in plasmin and complementary sites in alpha2-antiplasmin and in fibrinogen.

Plasminogen and plasminogen derivatives which contain lysine-binding sites were found to decrease the reaction rate between plasmin and alpha2-antiplasmin by competing with plasmin for the complementary site(s) in alpha2-antiplasmin. The dissocwation constant Kd for the interaction between intact plasminogen (Glu-plasminogen) and alpha2-antiplasmin is 4.0 microM but those for Lys-plasminogen or TLCK-plasmin are about 10-fold lower indicating a stronger interaction. The lysine-binding site(s) which is situated in triple-loops 1--3 in the plasmin A-chain is mainly responsible for the interaction with alpha2-antiplasmin. The interaction between Glu-plasminogen and alpha2-antiplasmin furthermore enhances the activation of Glu-plasminogen by urokinase to a comparable extent as 6-aminohexanoic acid, suggesting that similar conformational changes occur in the proenzyme after complex formation. Fibrinogen, fibrinogen digested with plasmin, purified fragment E and purified fragment D interfere with the reaction between plasmin and alpha2-antiplasmin by competing with alpha2-antiplasmin for the lysine-binding site(s) in the plasmin A-chain. The Kd obtained for these interactions varied between 0.2 microM and 1.4 microM; fragment E being the most effective. Thus the fibrinogen molecule contains several complementary sites to the lysine-binding sites located both in its NH2-terminal and COOH-terminal regions; these sites are to a large extent.

Binding Sites↗
PubMed1979

Turnover of antiplasmin, the fast-acting plasmin inhibitor of plasma.

Human antiplasmin, the fast-acting plasmin inhibitor in plasma, was purified to homogeneity and labeled with 125I. This material, which was indistinguishable from antiplasmin in plasma with respect to several physicochemical and functional properties, was injected intravenously and its turnover measured in control subjects and in patients undergoing thrombolytic therapy. In eight control subjects (four healthy persons and four patients with atherosclerosis), the following turnover characteristics were obtained: plasma radioactivity half-life 2.64 +/- 0.32 days, fractional catabolic rate 0.53 +/- 0.09 of the plasma pool per day, intravascular fraction 0.51 +/- 0.05, and synthetic (catabolic) rate 1.4 +/- 0.27 mg/kg/day. The half-life of the plasmin-antiplasmin complex in plasma, measured from the disappearance rate of labeled antiplasmin, plasmin or plasmin-antiplasmin complex during thrombolytic therapy was approximately 0.5 days.

Adult↗
PubMed1978

On the kinetics of the reaction between human antiplasmin and a low-molecular-weight form of plasmin.

The reaction between antiplasmin (A) and a low-molecular-weight form of plasmin (P) proceeds in at least two steps: a fast reversible second-order reaction followed by a slower irreversible first-order transition, and may be represented by: P +A k1 in equilibrium k-1 PA k2 leads to PA'. The low-Mr plasmin, which is obtained by limited elastase digestion, is composed of an intact B chain and a small A chain lacking the lysine-binding sites. The k1 of the reaction is (6.5 +/- 0.5) x 10(5) M-1 s-1 which is 30--60 times smaller than that for normal plasmin and antiplasmin. The dissociation constant of the first step is 1.9 x 10(-9) M which is 10 times higher than for normal plasmin and antiplasmin. The rate constant of the second step is (4.2 +/- 0.2) x 10(-3) s-1 for both normal and low-Mr plasmin. Low Mr plasmin which has substrate bound to its active site does not react or reacts only very slowly with antiplasmin. The reaction rate, however, is only slightly influenced by 6-aminohexanoic acid in concentrations up to 1 mM which decrease the reaction rate of normal plasmin approximately 50-fold. The findings further indicate that the lysine-binding site(s) of plasmin are of great importance for the rate of its reaction with antiplasmin.

Binding Sites↗
PubMed1978

Fast-acting plasmin inhibitor in human plasma.

The fast-acting and physiologically most important inhibitor of plasmin in human plasma is a recently discovered and purified alpha 2-glycoprotein with a molecular weight of 65,000-70,000 daltons occurring at a concentration of 1 muM. The inhibitor rapidly forms a completely inactive 1:1 stoichometric complex with plasmin through reaction with the B chain (light chain) of the enzyme, which contains the active center. It also reacts with trypsin and very slowly with urokinase and with some other enzymes in purified systems, but its role in vivo as an inhibitor of proteases other than plasmin seems negligible. Antiplasmin is the only plasma protein that can inhibit the fibrinolysis associated with transformed or malignant cells. The plasmin-antiplasmin complex contains neoantigenic structures not present in the parent molecules that may form the basis of immunochemical methods for detecting activation of the fibrinolvtic system in blood.

Cell Transformation, Neoplastic↗
PubMed1977

Primary structure of the B-chain of human plasmin.

The primary structure of the human plasmin B-chain has been determined. It consists of 230 residues divided in three cyanogen bromide fragments: The amino-terminal 24 residues, the carboxy-terminal three residues and the middle 203 residues. Sequence detemination was performed on the tryptic and the chymotryptic peptides obtained from the main cyanogen bromide fragment of this chain. Owing to similarities between some of the overlapping chymotryptic peptides, two different sequences were possible from these results. However, since the homologies with the pancreatic serine proteases and also the B-chains of thrombin and factor XA are pronounced, the arrangement still could be settled. By peptic digestion of partially reduced and S-carboxymethylated B-chain it was shown that there are two interchain disulphide bridges, which connect the A and B-chains of plasmin, involving Cys-5 and Cys-105 from the B-chain. The intrachain disulphides in the B-chain seem to be situated exactly as in chymotrypsin as partly judged from homologies.

Amino Acid Sequence↗