Urokinase-type plasminogen activator-like proteases in teleosts lack genuine receptor-binding epidermal growth factor-like domains.
Plasminogen activation catalyzed by urokinase-type plasminogen activator (uPA) plays an important role in normal and pathological tissue remodeling processes. Since its discovery in the mid-1980s, the cell membrane-anchored urokinase-type plasminogen activator receptor (uPAR) has been believed to be central to the functions of uPA, as uPA-catalyzed plasminogen activation activity appeared to be confined to cell surfaces through the binding of uPA to uPAR. However, a functional uPAR has so far only been identified in mammals. We have now cloned, recombinantly produced, and characterized two zebrafish proteases, zfuPA-a and zfuPA-b, which by several criteria are the fish orthologs of mammalian uPA. Thus, both proteases catalyze the activation of fish plasminogen efficiently and both proteases are inhibited rapidly by plasminogen activator inhibitor-1 (PAI-1). But zfuPA-a differs from mammalian uPA by lacking the exon encoding the uPAR-binding epidermal growth factor-like domain; zfuPA-b differs from mammalian uPA by lacking two cysteines of the epidermal growth factor-like domain and a uPAR-binding sequence comparable with that found in mammalian uPA. Accordingly, no zfuPA-b binding activity could be found in fish white blood cells or fish cell lines. We therefore propose that the current consensus of uPA-catalyzed plasminogen activation taking place on cell surfaces, derived from observations with mammals, is too narrow. Fish uPAs appear incapable of receptor binding in the manner known from mammals and uPA-catalyzed plasminogen activation in fish may occur mainly in solution. Studies with nonmammalian vertebrate species are needed to obtain a comprehensive understanding of the mechanism of plasminogen activation.
Department of Molecular Biology and Genetics, Aarhus University, 10 Gustav Wieds Vej, 8000 Aarhus C, Denmark.
René
R
Bager
Thomas K
TK
Kristensen
Jan K
JK
Jensen
Agnieszka
A
Szczur
Anni
A
Christensen
Lisbeth M
LM
Andersen
Jesper S
JS
Johansen
Niels
N
Larsen
Erik
E
Baatrup
Mingdong
M
Huang
Michael
M
Ploug
Peter A
PA
Andreasen
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D010958:Plasminogen / Q000235:genetics / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
D017395:Plasminogen Activator Inhibitor 1; D029961:Zebrafish Proteins; D010958:Plasminogen; D014568:Urokinase-Type Plasminogen Activator