Buffer interference with protein dynamics: a case study on human liver fatty acid binding protein.
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.
Long
Dong
D
Department of Biological Sciences, National University of Singapore, Singapore 117543.
Yang
Daiwen
D
D017738:Alkanesulfonic Acids / Q000737:chemistry*; D002021:Buffers; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D009025:Morpholines / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D014325:Tromethamine / Q000031:analogs & derivatives* / Q000737:chemistry
D017738:Alkanesulfonic Acids; D002021:Buffers; D050556:Fatty Acid-Binding Proteins; D009025:Morpholines; D014325:Tromethamine; C004550:2-(N-morpholino)ethanesulfonic acid; C026272:Bistris