Abstract
Protein molecules naturally emit streams of information-rich signals in the language of hydrogen exchange concerning the intimate details of their stability, dynamics, function, changes therein, and effects thereon, all resolved to the level of their individual amino acids. The effort to measure protein hydrogen exchange behavior, understand the underlying chemistry and structural physics of hydrogen exchange processes, and use this information to learn about protein properties and function has continued for 50 years. Recent work uses mass spectrometric analysis together with an earlier proteolytic fragmentation method to extend the hydrogen exchange capability to large biologically interesting proteins. This article briefly reviews the advances that have led us to this point and the understanding that has so far been achieved.
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Hvidt, A.; Nielsen, S. O. Hydrogen exchange in proteins. Adv. Protein Chem. 1966, 21, 287–386.
Eigen, M. Proton transfer, acid-base catalysis, and enzymatic hydrolysis. Angew. Chem. Int. Ed. English 1964, 3, 1–19.
Englander, S. W.; Kallenbach, N. R. Hydrogen exchange and structural dynamics of proteins and nucleic-acids. Q. Rev. Biophys. 1984, 16, 521–655.
Perrin, C. L. Proton exchange in amides: Surprises from simple systems. Accts. Chem. Res. 1989, 22, 268–275.
Bai, Y.; Milne, J. S.; Mayne, L.; Englander, S. W. Primary structure effects on peptide group hydrogen exchange. Proteins 1993, 17, 75–86.
Connelly, G. P.; Bai, Y.; Jeng, M. F.; Englander, S. W. Isotope effects in peptide group hydrogen exchange. Proteins 1993, 17, 87–92.
Hvidt, A. A discussion of the pH dependence of the hydrogen-deuterium exchange of proteins. C. R. Trav. Lab. Carlsberg 1964, 34, 299–317.
Krishna, M. M. G.; Hoang, L.; Lin, Y.; Englander, S. W. Hydrogen exchange methods to study protein folding. Methods 2004, 34, 51–64.
Rose, M. C.; Stuehr, J. Kinetics of proton transfer reactions in aqueous solution: rates of internally hydrogen bonded systems. J. Am. Chem. Soc. 1968, 90, 7205–7209.
Haslam, J. L.; Eyring, E. M. Deuterium oxide solvent isotope effects on N-H...O, O-H...N, and N-H.N intramolecular hydrogen bonds. J. Phys. Chem. 1967, 71, 4470–4475.
Milne, J. S.; Mayne, L.; Roder, H.; Wand, A. J.; Englander, S. W. Determinants of protein hydrogen exchange studied in equine cytochrome. c. Protein Sci. 1998, 7, 739–745.
Milne, J. S.; Xu, Y.; Mayne, L. C.; Englander, S. W. Experimental study of the protein folding landscape: Unfolding reactions in cytochrome. c. J. Mol. Biol. 1999, 290, 811–822.
Calhoun, D. B.; Vanderkooi, J. M.; Englander, S. W. Penetration of small molecules into proteins studied by quenching of phosphorescence and fluorescence. Biochemistry 1983, 22, 1533–1539.
Calhoun, D. B.; Englander, S. W.; Wright, W. W.; Vanderkooi, J. M. Quenching of room temperature protein phosphorescence by added small molecules. Biochemistry 1988, 27, 8466–8474.
Bai, Y.; Milne, J. S.; Mayne, L.; Englander, S. W. Protein stability parameters measured by hydrogen exchange. Proteins 1994, 20, 4–14.
Huyghues-Despointes, B. M. P.; Scholtz, J. M.; Pace, C. N. Protein conformational stabilities can be determined from hydrogen exchange rates. Nat. Struct. Biol. 1999, 6, 910–912.
Bai, Y.; Englander, S. W. Future directions in folding: The multi-state nature of protein structure. Proteins 1996, 24, 145–151.
Maity, H.; Lim, W. K.; Rumbley, J. N.; Englander, S. W. Protein hydrogen exchange mechanism: Local fluctuations. Protein Sci. 2003, 12, 153–160.
Englander, S. W. A hydrogen exchange method using tritium and Sephadex: Application to ribonuclease. Biochemistry 1963, 2, 798–807.
Perutz, M. F. Mechanisms of cooperativity and allosteric regulation in proteins. Q Rev Biophys 1989, 22, 139–237.
Englander, J. J.; Rogero, J. R.; Englander, S. W. Protein-hydrogen exchange studied by a fragment separation method. Anal. Biochem. 1985, 147, 234–244.
Rogero, J. R.; Englander, J. J.; Englander, S. W. Individual breathing reactions measured by functional labeling and hydrogen exchange methods. Methods Enzymol. 1986, 131, 508–517.
Englander, S. W.; Englander, J. J. Structure and energy change in hemoglobin by hydrogen exchange labeling. Methods Enzymol. 1994, 232, 26–42.
Wyman, J. Linked functions and reciprocal effects in hemoglobin: A second look. Adv. Protein Chem. 1964, 19, 223–286.
Monod, J.; Wyman, J.; Changeaux, J. P. On the nature of allosteric transitions: A plausible model. J. Mol. Biol. 1965, 12, 88–118.
Zhang, Z.; Smith, D. L. Determination of amide-hydrogen exchange by mass spectrometry: A new tool for protein structure elucidation. Protein Sci. 1993, 2, 522–531.
Kaltashov, I. A.; Eyles, S. J. Mass Spectrometry in Biophysics: Conformation and Dynamics of Biomolecules; John Wiley: Hoboken, NJ, 2005, p 458.
Hoerner, J. K.; Xiao, H.; Dobo, A.; Kaltashov, I. A. Is there hydrogen scrambling in the gas phase?: Energetic and structural determinants of proton mobility within ions. J. Am. Chem. Soc. 2004, 126, 7709–7717.
Delmar, C.; Greenbaum, E. A.; Mayne, L.; Englander, S. W.; Woods V. L. Jr. Structure and properties of α-synuclein and other amyloids determined at the amino acid level. Proc. Natl. Acad. Sci. U.S.A. 2005, 102, 15477–15482.
Hotchko, M.; Anand, G. S.; Komives, E. A.; Ten Eyck, L. F. Automated extraction of backbone deuteration levels from amide H/2H mass spectrometry experiments. Protein Sci. 2006, 15, 583–601.
Engen, J. R.; Smith, D. L. Investigating the higher order structure of proteins: Hydrogen exchange, proteolytic fragmentation, and mass spectrometry. Methods Mol. Biol. 2000, 146, 95–112.
Eyles, S. J.; Kaltashov, I. A. Methods to study protein dynamics and folding by mass spectrometry. Methods 2004, 34, 88–99.
Hoofnagle, A. N.; Resing, K. A.; Ahn, N. G. Protein analysis by hydrogen exchange mass spectrometry. Annu. Rev. Biophys. Biomol. Struct. 2003, 32, 1–25.
Wales, T. E.; Engen, J. R. Hydrogen exchange mass spectrometry for the analysis of protein dynamics. Mass Spectrom. Rev. 2006, 25, 158–170.
Englander, J. J.; Del Mar, C.; Li, W.; Englander, S. W.; Kim, J. S.; Stranz, D. D.; Hamuro, Y.; Woods V. C. Jr. Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry. Proc. Natl. Acad. Sci. U.S.A. 2003, 100, 7057–7062.
Englander, S. W.; Mauel, C. Hydrogen exchange studies of respiratory proteins: Detection of discrete ligand induced changes in hemoglobin. J. Biol. Chem. 1972, 247, 2387–2394.
Liem, R. K. H.; Calhoun, D. B.; Englander, J. J.; Englander, S. W. A high energy structure change in hemoglobin studied by difference hydrogen exchange. J. Biol. Chem. 1980, 255, 10687–10694.
Englander, J. J.; Rumbley, J. N.; Englander, S. W. Signal transmission between subunits in the hemoglobin T-state. J. Mol. Biol. 1998, 284, 1707–1716.
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Published online July 28, 2006
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Englander, S.W. Hydrogen exchange and mass spectrometry: A historical perspective. The official journal of The American Society for Mass Spectrometry 17, 1481–1489 (2006). https://doi.org/10.1016/j.jasms.2006.06.006
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DOI: https://doi.org/10.1016/j.jasms.2006.06.006