Structure and function of microplasminogen: II. Determinants of activation by urokinase and by the bacterial activator streptokinase.
We have used a group of human microplasminogens (mPlg), modified by residue substitutions, insertions, deletions, and chain breaks (1) to study the determinants of productive interactions with two plasminogen activators, urokinase (uPA), and streptokinase (SK); (2) to explore the basis of species specificity in the zymogen-SK complex activity; and (3) to compare active SK complex formation in mPlg and microplasmin (mPlm). Modifications within the disulfide-bonded loop containing the activation site and the adjacent hexadecapeptide upstream sequence showed that uPA recognition elements encompassed R29 at the activation site and multiple elements extending upstream to perhaps 13 residues, all maintained in specific conformational register by surrounding pairs of disulfide bonds. A generally parallel pattern of structural requirements was observed for active zymogen-SK complex formation. Changes within the loop downstream of the activation site were tolerated well by uPA and poorly by SK. The introduction of selected short bovine (Plg) sequences in human mPlg reduced the activity of the resulting SK complexes. The requirements for active SK complex formation are different for mPlg and mPlm.
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794, USA.
J
J
Wang
E
E
Reich
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D004220:Disulfides / Q000737:chemistry; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010958:Plasminogen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D011994:Recombinant Proteins; D013045:Species Specificity; D013300:Streptokinase / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D004220:Disulfides; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D010958:Plasminogen; D013300:Streptokinase; D014568:Urokinase-Type Plasminogen Activator