English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
 
 
DownloadE-Mail
  Structural analysis of the mitotic regulator hPin1 in solution - Insights into domain architecture and substrate binding

Bayer, E., Goettsch, S., Mueller, J. W., Griewel, B., Guiberman, E., Mayr, L. M., et al. (2003). Structural analysis of the mitotic regulator hPin1 in solution - Insights into domain architecture and substrate binding. Journal of Biological Chemistry, 278(28), 26183-26193. Retrieved from http://www.jbc.org/cgi/content/abstract/278/28/26183.

Item is

Basic

show hide
Genre: Journal Article
Alternative Title : J. Biol. Chem.

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Bayer, Elena1, Author
Goettsch, Sandra1, Author
Mueller, Jonathan W.1, Author
Griewel, Bernhard1, Author
Guiberman, Elena1, Author
Mayr, Lorenz M., Author
Bayer, Peter2, Author           
Affiliations:
1Max Planck Institute of Molecular Physiology, Max Planck Society, ou_1753286              
2Sonstige Wissenschaftliche Organisationseinheiten, Max Planck Institute of Molecular Physiology, Max Planck Society, ou_1753294              

Content

show
hide
Free keywords: PROLYL ISOMERASE PIN1; DEPENDENT PROLINE ISOMERIZATION; PHOSPHOSERINE-BINDING; PHOSPHORYLATION; MITOSIS; P53; RECOGNITION
 Abstract: The peptidyl-prolyl cis/trans isomerase hPin1 is a phosphorylation-dependent regulatory enzyme whose substrates are proteins involved in regulation of cell cycle, transcription, Alzheimer's disease, and cancer pathogenesis. We have determined the solution structure of the two domain protein hPin1-(1-163) and its separately expressed PPIase domain ( 50-163) ( hPin1(PPIase)) with an root mean square deviation of <0.5 &ANGS; over backbone atoms using NMR. Domain organization of hPin1 differs from that observed in structures solved by x-ray crystallography. Whereas PPIase and WW domain are tightly packed onto each other and share a common binding interface in crystals, our NMR-based data revealed only weak interaction of both domains at their interface in solution. Interaction between the two domains of full-length hPin1 is absent when the protein is dissected into the catalytic and the WW domain. It indicates that the flexible linker, connecting both domains, promotes binding. By evaluation of NOESY spectra we can show that the α1/β1 loop, which was proposed to undergo a large conformational rearrangement in the absence of sulfate and an Ala-Pro peptide, remained in the closed conformation under these conditions. Dissociation constants of 0.4 and 2.0 mM for sulfate and phosphate ions were measured at 12 &DEG;C by fluorescence spectroscopy. Binding of sulfate prevents hPin1 aggregation and changes surface charges across the active center and around the reactive and catalytically essentialCys(113). In the absence of sulfate and/or reducing agent this residue seems to promote aggregation, as observed in hPin1 solutions in vitro.

Details

show
hide
Language(s): eng - English
 Dates: 2003-07-11
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of Biological Chemistry
  Alternative Title : J. Biol. Chem.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 278 (28) Sequence Number: - Start / End Page: 26183 - 26193 Identifier: -