Substitution of arginine 719 for glutamic acid in human plasminogen substantially reduces its affinity for streptokinase.
In isolation human plasminogen possesses no enzymatic activity, yet upon formation of an equimolar complex with the bacterial protein streptokinase, it acquires a plasminogen activator function. The region(s) of plasminogen and of streptokinase which mediate complex formation has (have) not been previously published. Here it is reported that a single-residue substitution (Arg719-->Glu) in the serine protease domain of full-length Glu-plasminogen substantially reduces its affinity for streptokinase. The plasminogen variant displays no other significant differences from the wild-type molecule with respect to activation by two-chain urokinase-type plasminogen activator, recognition by monoclonal antibodies, or ability to undergo conformational change. It is concluded that Arg719 in human plasminogen is an important determinant of the streptokinase binding site, although further sites are likely to contribute both to the affinity of plasminogen for streptokinase and to mechanisms by which the active site is formed within the complex.
Dawson
K M
KM
British Biotech, Cowley, Oxford, U.K.
Marshall
J M
JM
Raper
R H
RH
Gilbert
R J
RJ
Ponting
C P
CP
D000818:Animals; D000911:Antibodies, Monoclonal; D001120:Arginine / Q000737:chemistry*; D001483:Base Sequence; D001665:Binding Sites; D016466:CHO Cells; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D018698:Glutamic Acid / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides / Q000737:chemistry; D010958:Plasminogen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010960:Plasminogen Activators; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013300:Streptokinase / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D009841:Oligonucleotides; D011994:Recombinant Proteins; D018698:Glutamic Acid; D010958:Plasminogen; D001120:Arginine; D013300:Streptokinase; D010960:Plasminogen Activators; D012697:Serine Endopeptidases