3PB1

Crystal Structure of a Michaelis Complex between Plasminogen Activator Inhibitor-1 and Urokinase-type Plasminogen Activator

Summary for 3PB1

• Entry DOI: 10.2210/pdb3pb1/pdb
• Related: 1DVM 2O8T
• Descriptor: Plasminogen activator inhibitor 1, Plasminogen activator, urokinase, SULFATE ION, ... (4 entities in total)
• Functional Keywords: pai-1, upa, michaelis complex, structural genomics, structure 2 function project, s2f, hydrolase inhibitor-hydrolase complex, hydrolase inhibitor/hydrolase
• Biological source: Homo sapiens (human); More
• Cellular location: Secreted: P05121
• Total number of polymer chains: 2
• Total formula weight: 71413.57

Authors

Lin, Z.,Jiang, L.,Huang, M.,Structure 2 Function Project (S2F) (deposition date: 2010-10-20, release date: 2010-12-29, Last modification date: 2024-11-06)

Primary citation

Lin, Z.,Jiang, L.,Yuan, C.,Jensen, J.K.,Zhang, X.,Luo, Z.,Furie, B.C.,Furie, B.,Andreasen, P.A.,Huang, M.
Structural basis for recognition of urokinase-type plasminogen activator by plasminogen activator inhibitor-1.
J.Biol.Chem., 286:7027-7032, 2011

PubMed Abstract: Plasminogen activator inhibitor-1 (PAI-1), together with its physiological target urokinase-type plasminogen activator (uPA), plays a pivotal role in fibrinolysis, cell migration, and tissue remodeling and is currently recognized as being among the most extensively validated biological prognostic factors in several cancer types. PAI-1 specifically and rapidly inhibits uPA and tissue-type PA (tPA). Despite extensive structural/functional studies on these two reactions, the underlying structural mechanism has remained unknown due to the technical difficulties of obtaining the relevant structures. Here, we report a strategy to generate a PAI-1·uPA(S195A) Michaelis complex and present its crystal structure at 2.3-Å resolution. In this structure, the PAI-1 reactive center loop serves as a bait to attract uPA onto the top of the PAI-1 molecule. The P4-P3' residues of the reactive center loop interact extensively with the uPA catalytic site, accounting for about two-thirds of the total contact area. Besides the active site, almost all uPA exosite loops, including the 37-, 60-, 97-, 147-, and 217-loops, are involved in the interaction with PAI-1. The uPA 37-loop makes an extensive interaction with PAI-1 β-sheet B, and the 147-loop directly contacts PAI-1 β-sheet C. Both loops are important for initial Michaelis complex formation. This study lays down a foundation for understanding the specificity of PAI-1 for uPA and tPA and provides a structural basis for further functional studies.

PubMed: 21199867
DOI: 10.1074/jbc.M110.204537

Experimental method

X-RAY DIFFRACTION (2.3 Å)