Two distinct pathways of the streptokinase-mediated activation of highly purified human plasminogen

McClintock, David K.; Englert, Mary E.; Dziobkowski, C.; Snedeker, E. H.; Bell, Paul H.

doi:10.1021/bi00723a013

Cited by 37 publications

(19 citation statements)

References 50 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Deletion of residues 1-59 (rSK⌬59) did not affect the K m grossly but markedly reduced the k cat (Ͼ600-fold decrease relative to rSK), effectively eliminating Pg activation. This finding indicates that the rSK1-59 peptide, particularly residues 24-59, is critical for Pg activation and that the peptide plays a more important role than is generally appreciated (27)(28)(29)(30)(31).…”

Section: Resultsmentioning

confidence: 89%

“…Beyond these domains, the NH 2 -and COOH-terminal residues were found to be disordered. During Pg activation in vitro or in human plasma, SK is cleaved within seconds to minutes at its NH 2 peptide plays a role in Pg activation, although there are significant disagreements about the magnitude of the peptide's effects and the stoichiometry between it and the core fragment (27)(28)(29)(30)(31). expressed as a factor Xaa-cleavable fusion protein in bacteria via the pMALc vector (New England Biolabs), cleaved, and purified as described in detail (33).…”

mentioning

confidence: 99%

See 1 more Smart Citation

A catalytic switch and the conversion of streptokinase to a fibrin-targeted plasminogen activator

Reed

Houng

Liu

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Plasminogen (Pg) activators such as streptokinase (SK) save lives by generating plasmin to dissolve blood clots. Some believe that the unique ability of SK to activate Pg in the absence of fibrin limits its therapeutic utility. We have found that SK contains an unusual NH 2 -terminal ''catalytic switch'' that allows Pg activation through both fibrin-independent and fibrin-dependent mechanisms. Unlike SK, a mutant (rSK⌬59) fusion protein lacking the 59 NH 2 -terminal residues was no longer capable of fibrinindependent Pg activation (k cat ͞K m decreased by >600-fold). This activity was restored by coincubation with equimolar amounts of the NH 2 -terminal peptide rSK1-59. Deletion of the NH 2 terminus made rSK⌬59 a Pg activator that requires fibrin, but not fibrinogen, for efficient catalytic function. The fibrin-dependence of the rSK⌬59 activator complex apparently resulted from selective catalytic processing of fibrinbound Pg substrates in preference to other Pg forms. Consistent with these observations, the presence (rSK) or absence (rSK⌬59) of the SK NH 2 -terminal peptide markedly altered fibrinolysis of human clots suspended in plasma. Like native SK, rSK produced incomplete clot lysis and complete destruction of plasma fibrinogen; in contrast, rSK⌬59 produced total clot lysis and minimal fibrinogen degradation. These studies indicate that structural elements in the NH 2 terminus are responsible for SK's unique mechanism of fibrin-independent Pg activation. Because deletion of the NH 2 terminus alters SK's mechanism of action and targets Pg activation to fibrin, there is the potential to improve SK's therapeutic efficacy.

show abstract

Section: Resultsmentioning

confidence: 89%

mentioning

confidence: 99%

A catalytic switch and the conversion of streptokinase to a fibrin-targeted plasminogen activator

Reed

Houng

Liu

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Human plasminogen was prepared by affinity chromatography on L-lysine-Sepharose by the method of Deutsch and Mertz [5] and McClintock et al [20] without separation of the different forms [21,22] of Lys-plasminogen and Glu-plasminogen. Lys-plasminogen was isolated from frozen human serum Cohn fraction 111 obtained from the American Red Cross.…”

Section: Enzymes and Chemicalsmentioning

confidence: 99%

Studies on the Lysine-Binding Sites of Human Plasminogen. The Effect of Ligand Structure on the Binding of Lysine Analogs to Plasminogen

Winn

Hochschwender

et al. 1980

Eur J Biochem

View full text Add to dashboard Cite

A method is described for measuring relative binding constants of lysine and analogs of lysine to plasminogen and plasminogen 'kringle' fragments. Plasminogen or kringle fragments adsorbed to lysine-Sepharose are eluted with increasing concentrations of lysine or other ligands, the concentration of ligand required to elute 50% of the protein being taken as a measure of the binding constant. The method is simple and is not dependent on monitoring conformational changes. We confirm earlier reports that the best ligands for the lysine binding sites of plasminogen are w-amino acids containing five or six carbons. We show further that both Glu-plasminogen (the native form with N-terminal glutamic acid) and Lys-plasminogen (a degraded form with N-terminal lysine), as well as the heavy chain fragments, kringle 4 and kringle 1 + 2 + 3, have very similar properties with regard to binding specificity for w-amino acids. For all species optimal binding is observed when the distance between the amino and carboxyl carbon is about 0.68 nm. The binding of ligands is decreased by the presence of polar atoms on the a and p positions of the carbon chain of amino acids.Arginine binds relatively weakly at the lysine site and there does not appear to be a separate arginine binding site in plasminogen.It has been known for over 20 years that amino acids such as 6-aminohexanoic acid have antifibrinolytic properties [l], one of which probably results from inhibition of the binding of plasminogen to fibrin [2]. Subsequent studies have shown that 6-aminohexanoic acid and lysine cause extensive conformational alterations in Glu-plasminogen [3,4] (and references in [4]); Deutsch and Mertz [5] utilized the strong affinity of plasminogen for lysine-Sepharose in the development of a convenient method for purification of plasminogen.

show abstract

“…Human plasminogen (lysine form [7]) was prepared from Cohn fraction III or IIIZ,a by the method of McClintock et al [8].…”

Section: Methodsmentioning

confidence: 99%

The primary structure of human plasminogen: Characterization and alignment of the cyanogen bromide peptides

Lee

Laursen

1976

FEBS Letters

View full text Add to dashboard Cite

Two distinct pathways of the streptokinase-mediated activation of highly purified human plasminogen

Cited by 37 publications

References 50 publications

A catalytic switch and the conversion of streptokinase to a fibrin-targeted plasminogen activator

A catalytic switch and the conversion of streptokinase to a fibrin-targeted plasminogen activator

Studies on the Lysine-Binding Sites of Human Plasminogen. The Effect of Ligand Structure on the Binding of Lysine Analogs to Plasminogen

The primary structure of human plasminogen: Characterization and alignment of the cyanogen bromide peptides

Contact Info

Product

Resources

About