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  • Buffer interference with protein dynamics: a case study on human liver fatty acid binding protein.

Buffer interference with protein dynamics: a case study on human liver fatty acid binding protein.

Biophysical journal (2009-02-17)
Dong Long, Daiwen Yang
ABSTRACT

Selection of suitable buffer types is often a crucial step for generating appropriate protein samples for NMR and x-ray crystallographic studies. Although the possible interaction between MES buffer (2-(N-morpholino)ethanesulfonic acid) and proteins has been discussed previously, the interaction is usually thought to have no significant effects on the structures of proteins. In this study, we demonstrate the direct, albeit weak, interaction between MES and human liver fatty acid binding protein (hLFABP). Rather than affecting the structure of hLFABP, we found that the dynamics of hLFABP, which were previously proposed to be relevant to its functions, were significantly affected by the binding of hLFABP with MES. Buffer interference with protein dynamics was also demonstrated with Bis-Tris buffer, which is quite different from MES and fatty acids in terms of their molecular structures and properties. This result, to our knowledge, is the first published report on buffer interference with protein dynamics on a microsecond to millisecond timescale and could represent a generic problem in the studies of functionally relevant protein dynamics. Although being a fortuity, our finding of buffer-induced changes in protein dynamics offers a clue to how hLFABP accommodates its ligands.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MES hydrate, BioUltra, ≥99.5% (T)
Sigma-Aldrich
MES hydrate, ≥99.5% (titration)
Sigma-Aldrich
MES hydrate, ≥99.5% (titration), pH 2.5-4.0 (0.5 M in H2O), BioXtra
Sigma-Aldrich
MES hydrate, BioPerformance Certified, suitable for cell culture, ≥99.5%
Sigma-Aldrich
MES, low moisture content, ≥99% (titration)
Sigma-Aldrich
MES sodium salt, BioPerformance Certified, suitable for cell culture
Sigma-Aldrich
BIS-TRIS, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, ≥98.0%
Sigma-Aldrich
BIS-TRIS, BioXtra, ≥98.0% (titration)
Sigma-Aldrich
BIS-TRIS, ≥98.0% (titration)
Sigma-Aldrich
MES sodium salt, ≥99% (titration)
Sigma-Aldrich
MES sodium salt, BioReagent, ≥99% (titration), crystalline
Sigma-Aldrich
BIS-TRIS, BioUltra, ≥99.0% (NT)
SAFC
MES sodium salt
SAFC
BIS-TRIS
Sigma-Aldrich
MES solution, BioUltra, for molecular biology, 0.5 M in H2O