GuHCl and NaCl-dependent hydrogen exchange in MerP reveals a well-defined core with an unusual exchange pattern.
We have analysed hydrogen exchange at amide groups to characterise the energy landscape of the 72 amino acid residue protein MerP. From the guanidine hydrochloride (GuHCl) dependence of exchange in the pre-transitional region we have determined free energy values of exchange (DeltaG(HX)) and corresponding m-values for individual amide protons. Detailed analysis of the exchange patterns indicates that for one set of amide protons there is a weak dependence on denaturant, indicating that the exchange is dominated by local fluctuations. For another set of amide protons a linear, but much stronger, denaturant dependence is observed. Notably, the plots of free energy of exchange versus [GuHCl] for 16 amide protons show pronounced upward curvature, and a close inspection of the structure shows that these residues form a well-defined core in the protein. The hydrogen exchange that was measured at various concentrations of NaCl shows an apparent selective stabilisation of this core. Detailed analysis of this exchange pattern indicates that it may originate from selective destabilisation of the unfolded state by guanidinium ions and/or selective stabilisation of the core in the native state by chloride ions.
Department of Biochemistry, Umeå University, SE-901 87 Umeå, Sweden.
Ann-Christin
AC
Brorsson
Martin
M
Lundqvist
Ingmar
I
Sethson
Bengt-Harald
BH
Jonsson
D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D019791:Guanidine / Q000737:chemistry* / Q000378:metabolism; D006859:Hydrogen / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010277:Parasympathomimetics / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D011522:Protons; D012965:Sodium Chloride / Q000737:chemistry*
D000577:Amides; C086060:MerP protein, Bacteria; D010277:Parasympathomimetics; D011506:Proteins; D011522:Protons; D012965:Sodium Chloride; D006859:Hydrogen; D019791:Guanidine