Combined use of NMR relaxation measurements and hydrodynamic calculations to study protein association. Evidence for tetramers of low molecular weight protein tyrosine phosphatase in solution
Bernado, P ; Åkerud, Tomas LU ; Garcia de la Torre, J ; Akke, Mikael LU
and Pons, M
(2003)
In Journal of the American Chemical Society
125(4).
p.916-923
- Abstract
- We describe a novel method for determining weak association constants of oligomeric protein complexes formed transiently under equilibrium conditions. This type of equilibrium process is recognized as being biologically important, but generally hard to study. Heteronuclear spin relaxation rates measured at multiple protein concentrations are analyzed using relaxation rates predicted from hydrodynamic calculations, yielding equilibrium constants and structural characterization of the protein complexes. The method was used to study the oligomerization equilibrium of bovine low molecular weight protein tyrosine phosphatase. X-ray structures of monomeric and dimeric forms of the protein have been reported previously. Using longitudinal and... (More)
- We describe a novel method for determining weak association constants of oligomeric protein complexes formed transiently under equilibrium conditions. This type of equilibrium process is recognized as being biologically important, but generally hard to study. Heteronuclear spin relaxation rates measured at multiple protein concentrations are analyzed using relaxation rates predicted from hydrodynamic calculations, yielding equilibrium constants and structural characterization of the protein complexes. The method was used to study the oligomerization equilibrium of bovine low molecular weight protein tyrosine phosphatase. X-ray structures of monomeric and dimeric forms of the protein have been reported previously. Using longitudinal and transverse 15N relaxation rates measured at four different protein concentrations, we detected the monomer, dimer, and a previously unknown tetramer and measured the dissociation constants of the equilibria involving these species. A comparison of experimental and predicted relaxation rates for individual backbone amide 15N spins enabled delineation of the tetramerization interface. The results suggest a novel concept for substrate modulation of enzymatic activity based on a "supramolecular proenzyme". The fast and reversible switching of the "supramolecular proenzyme" would have obvious advantages for the regulation of enzymes involved in cell signaling pathways. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/128023
- author
- Bernado, P
; Åkerud, Tomas
LU
; Garcia de la Torre, J
; Akke, Mikael
LU
and Pons, M
- organization
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of the American Chemical Society
- volume
- 125
- issue
- 4
- pages
- 916 - 923
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000180579600022
- pmid:12537489
- scopus:0346634883
- ISSN
- 1520-5126
- DOI
- 10.1021/ja027836h
- language
- English
- LU publication?
- yes
- id
- 3e6f84dc-7ead-4e00-a725-3640fefe1810 (old id 128023)
- date added to LUP
- 2016-04-01 16:21:00
- date last changed
- 2022-01-28 19:06:46
@article{3e6f84dc-7ead-4e00-a725-3640fefe1810,
abstract = {{We describe a novel method for determining weak association constants of oligomeric protein complexes formed transiently under equilibrium conditions. This type of equilibrium process is recognized as being biologically important, but generally hard to study. Heteronuclear spin relaxation rates measured at multiple protein concentrations are analyzed using relaxation rates predicted from hydrodynamic calculations, yielding equilibrium constants and structural characterization of the protein complexes. The method was used to study the oligomerization equilibrium of bovine low molecular weight protein tyrosine phosphatase. X-ray structures of monomeric and dimeric forms of the protein have been reported previously. Using longitudinal and transverse 15N relaxation rates measured at four different protein concentrations, we detected the monomer, dimer, and a previously unknown tetramer and measured the dissociation constants of the equilibria involving these species. A comparison of experimental and predicted relaxation rates for individual backbone amide 15N spins enabled delineation of the tetramerization interface. The results suggest a novel concept for substrate modulation of enzymatic activity based on a "supramolecular proenzyme". The fast and reversible switching of the "supramolecular proenzyme" would have obvious advantages for the regulation of enzymes involved in cell signaling pathways.}},
author = {{Bernado, P and Åkerud, Tomas and Garcia de la Torre, J and Akke, Mikael and Pons, M}},
issn = {{1520-5126}},
language = {{eng}},
number = {{4}},
pages = {{916--923}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Journal of the American Chemical Society}},
title = {{Combined use of NMR relaxation measurements and hydrodynamic calculations to study protein association. Evidence for tetramers of low molecular weight protein tyrosine phosphatase in solution}},
url = {{http://dx.doi.org/10.1021/ja027836h}},
doi = {{10.1021/ja027836h}},
volume = {{125}},
year = {{2003}},
}