Hydrogen exchange methods to study protein folding.
The measurement of amino acid-resolved hydrogen exchange (HX) has provided the most detailed information so far available on the structure and properties of protein folding intermediates. Direct HX measurements can define the structure of tenuous molten globule forms that are generally inaccessible to the usual crystallographic and NMR methods (C. Redfield review in this issue). HX pulse labeling methods can specify the structure, stability and kinetics of folding intermediates that exist for less than 1 s during kinetic folding. Native state HX methods can detect and characterize folding intermediates that exist as infinitesimally populated high energy excited state forms under native conditions. The results obtained in these ways suggest principles that appear to explain the properties of partially folded intermediates and how they are organized into folding pathways. The application of these methods is detailed here.
Krishna
Mallela M G
MM
Johnson Research Foundation, Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6059, USA. mmg@hx2.med.upenn.edu
Hoang
Linh
L
Lin
Yan
Y
Englander
S Walter
SW
D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry; D041961:Deuterium Exchange Measurement; D006859:Hydrogen / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013997:Time Factors
D003574:Cytochrome c Group; D006859:Hydrogen