title: Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry.
abstract: An automated high-throughput, high-resolution deuterium exchange HPLC-MS method (DXMS) was used to extend previous hydrogen exchange studies on the position and energetic role of regulatory structure changes in hemoglobin. The results match earlier highly accurate but much more limited tritium exchange results, extend the analysis to the entire sequence of both hemoglobin subunits, and identify some energetically important changes. Allosterically sensitive amide hydrogens located at near amino acid resolution help to confirm the reality of local unfolding reactions and their use to evaluate resolved structure changes in terms of allosteric free energy.
authors.affiliation: Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Joan J
authors.initials: JJ
authors.lastname: Englander
authors.forename: Charyl
authors.initials: C
authors.lastname: Del Mar
authors.forename: Will
authors.initials: W
authors.lastname: Li
authors.forename: S Walter
authors.initials: SW
authors.lastname: Englander
authors.forename: Jack S
authors.initials: JS
authors.lastname: Kim
authors.forename: David D
authors.initials: DD
authors.lastname: Stranz
authors.forename: Yoshitomo
authors.initials: Y
authors.lastname: Hamuro
authors.forename: Virgil L
authors.initials: VL
authors.lastname: Woods
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003903:Deuterium / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry; D006801:Humans; D006859:Hydrogen / Q000737:chemistry; D066298:In Vitro Techniques; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry; D011489:Protein Denaturation; D021122:Protein Subunits; D011506:Proteins / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D006454:Hemoglobins; D010446:Peptide Fragments; D021122:Protein Subunits; D011506:Proteins; D006859:Hydrogen; D003903:Deuterium