5BRR

Michaelis complex of tPA-S195A:PAI-1


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 3.16 Å
  • R-Value Free: 0.276 
  • R-Value Work: 0.203 
  • R-Value Observed: 0.207 

wwPDB Validation   3D Report Full Report


This is version 1.4 of the entry. See complete history


Literature

Crystal Structure of the Michaelis Complex between Tissue-type Plasminogen Activator and Plasminogen Activators Inhibitor-1

Gong, L.Liu, M.Zeng, T.Shi, X.Yuan, C.Andreasen, P.A.Huang, M.

(2015) J Biol Chem 290: 25795-25804

  • DOI: https://doi.org/10.1074/jbc.M115.677567
  • Primary Citation of Related Structures:  
    5BRR

  • PubMed Abstract: 

    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.


  • Organizational Affiliation

    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.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Plasminogen activator inhibitor 1A [auth I]379Homo sapiensMutation(s): 4 
Gene Names: SERPINE1PAI1PLANH1
UniProt & NIH Common Fund Data Resources
Find proteins for P05121 (Homo sapiens)
Explore P05121 
Go to UniProtKB:  P05121
PHAROS:  P05121
GTEx:  ENSG00000106366 
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP05121
Sequence Annotations
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  • Reference Sequence
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 2
MoleculeChains Sequence LengthOrganismDetailsImage
Tissue-type plasminogen activatorB [auth E]252Homo sapiensMutation(s): 3 
Gene Names: PLAT
EC: 3.4.21.68
UniProt & NIH Common Fund Data Resources
Find proteins for P00750 (Homo sapiens)
Explore P00750 
Go to UniProtKB:  P00750
PHAROS:  P00750
GTEx:  ENSG00000104368 
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP00750
Sequence Annotations
Expand
  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 3.16 Å
  • R-Value Free: 0.276 
  • R-Value Work: 0.203 
  • R-Value Observed: 0.207 
  • Space Group: P 21 21 21
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 49.74α = 90
b = 73.94β = 90
c = 201.8γ = 90
Software Package:
Software NamePurpose
REFMACrefinement
MOLREPmodel building
HKL-2000data collection

Structure Validation

View Full Validation Report



Entry History & Funding Information

Deposition Data

  • Released Date: 2015-09-02 
  • Deposition Author(s): Gong, L.

Funding OrganizationLocationGrant Number
National Natural Science Foundation of ChinaChina31170707
National Natural Science Foundation of ChinaChina31370737
CAS/SFEA International Partnership Program for Creative Research TeamsChina--

Revision History  (Full details and data files)

  • Version 1.0: 2015-09-02
    Type: Initial release
  • Version 1.1: 2015-09-09
    Changes: Database references
  • Version 1.2: 2015-09-16
    Changes: Database references
  • Version 1.3: 2015-11-11
    Changes: Database references
  • Version 1.4: 2017-10-18
    Changes: Author supporting evidence, Database references, Derived calculations