Crystal Structure of the Michaelis Complex between Tissue-type Plasminogen Activator and Plasminogen Activators Inhibitor-1.
Thrombosis is a leading cause of death worldwide. Recombinant tissue-type plasminogen activator (tPA) is the Food and Drug Administration-approved thrombolytic drug. tPA is rapidly inactivated by endogenous plasminogen activator inhibitor-1 (PAI-1). Engineering on tPA to reduce its inhibition by PAI-1 without compromising its thrombolytic effect is a continuous effort. Precise details, with atomic resolution, of the molecular interactions between tPA and PAI-1 remain unknown despite previous extensive studies. Here, we report the crystal structure of the tPA·PAI-1 Michaelis complex, which shows significant differences from the structure of its urokinase-type plasminogen activator analogue, the uPA·PAI-1 Michaelis complex. The PAI-1 reactive center loop adopts a unique kinked conformation. The structure provides detailed interactions between tPA 37- and 60-loops with PAI-1. On the tPA side, the S2 and S1β pockets open up to accommodate PAI-1. This study provides structural basis to understand the specificity of PAI-1 and to design newer generation of thrombolytic agents with reduced PAI-1 inactivation.
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China, the University of Chinese Academy of Sciences, Beijing, 100049, China, and.
Lihu
L
Gong
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China, the University of Chinese Academy of Sciences, Beijing, 100049, China, and.
Min
M
Liu
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China.
Tu
T
Zeng
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China.
Xiaoli
X
Shi
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China.
Cai
C
Yuan
the Danish-Chinese Centre for Proteases and Cancer, Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark.
Peter A
PA
Andreasen
From the State Key Laboratory of Structural Chemistry and Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, 350002 Fujian, China, the University of Chinese Academy of Sciences, Beijing, 100049, China, and mhuang@fjirsm.ac.cn.
Mingdong
M
Huang
D018360:Crystallography, X-Ray; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D015394:Molecular Structure; D017395:Plasminogen Activator Inhibitor 1 / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D010959:Tissue Plasminogen Activator / Q000737:chemistry*
D017395:Plasminogen Activator Inhibitor 1; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator
Michaelis complex; PAI-1; crystal structure; fibrinolysis; serine protease; serpin; structural biology; tPA; thrombolytic agents