PMID: 2496749Jan 24, 1989Paper

The single-chain form of tissue-type plasminogen activator has catalytic activity: studies with a mutant enzyme that lacks the cleavage site

Biochemistry
J A BooseM J Gething

Abstract

Tissue-type plasminogen activator (t-PA), the serine protease responsible for catalyzing the production of plasmin from plasminogen at the site of blood clots, is synthesized as a single-chain polypeptide precursor. Proteolytic cleavage at the C-terminal side of Arg275 generates a two-chain form of the enzyme whose subunits are held together by a single disulfide bond. We have measured the activities of both forms of the wild-type enzyme, as well as that of a mutant enzyme (Arg275----Gly), created by oligonucleotide-directed mutagenesis, that cannot be cleaved into a two-chain form. Both types of single-chain t-PAs are enzymatically active and exhibit identical Vmax and Km values when assayed with synthetic peptide substrates, indicating that the single amino acid change had no effect on the amidolytic activity of the enzyme. However, cleavage of wild-type t-PA into the two-chain form results in increased activity both on a peptide substrate and on the natural substrates Lys- and Glu-plasminogen in the absence or presence of stimulation by soluble fibrin. The enhanced activity is due to a 3-5-fold increase in the Vmax of the cleaved enzyme, rather than to any change in the Km values for the various substrates. During incubation...Continue Reading

Citations

Oct 1, 1991·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·G LeloupG Vaes
Jun 4, 1993·Biochimica Et Biophysica Acta·S S MargossianJ McDonagh
Mar 15, 1995·Biochimica Et Biophysica Acta·K KolevR Machovich
Aug 9, 2012·The Journal of Biological Chemistry·Julie A ZornJames A Wells
Jun 23, 2001·Protein Science : a Publication of the Protein Society·A PasternakL Hedstrom
Aug 12, 2009·American Journal of Physiology. Lung Cellular and Molecular Physiology·Andrey A KomissarovSteven Idell
Jun 24, 1992·Biochimica Et Biophysica Acta·D M MillerP M Horowitz
Nov 4, 2015·Frontiers in Cellular Neuroscience·Arnaud ChevilleyDenis Vivien
Apr 21, 2005·Protein Science : a Publication of the Protein Society·Henry R MaunRobert A Lazarus
Jul 22, 2014·Journal of Thrombosis and Haemostasis : JTH·R EngelN J Mutch
Nov 18, 2014·Molecular Microbiology·Ulrike SiegmundPaul Tudzynski
Sep 29, 2012·The Biochemical Journal·Longguang JiangMingdong Huang
Jul 6, 2016·Frontiers in Cellular Neuroscience·Karen-Sue B CarlsonBradford S Schwartz
Oct 16, 1990·Biochemistry·M G ObukowiczD T McPherson
Jan 3, 1997·The Journal of Biological Chemistry·K Tachias, E L Madison
Jul 26, 2017·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Romain GoulayJérôme Parcq
Apr 21, 2018·PLoS Neglected Tropical Diseases·Adrian LeontovyčMartin Horn
Nov 15, 1996·The Journal of Biological Chemistry·K Tachias, E L Madison
Jun 26, 2004·The Journal of Biological Chemistry·Daniel KirchhoferRobert A Lazarus
Jun 29, 1989·Nature·E L MadisonJ F Sambrook
May 3, 2002·European Journal of Biochemistry·Inez Ruiz-StewartScott A Waldman
Aug 4, 1995·The Journal of Biological Chemistry·K Tachias, E L Madison
Sep 14, 2002·The Journal of Biological Chemistry·Karen Barker-CarlsonBradford S Schwartz
Mar 11, 2020·Experimental & Molecular Medicine·Haili LinPeng Xu
Oct 27, 1995·The Journal of Biological Chemistry·P BringmannP Donner

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