Identifying Zn-bound histidine residues in metalloproteins using hydrogen-deuterium exchange mass spectrometry.

Abstract

In this work, we have developed a method that uses hydrogen-deuterium exchange (HDX) of C2-hydrogens of histidines coupled with mass spectrometry (MS) to identify Zn-bound histidines in metalloproteins. This method relies on differences in HDX reaction rates of Zn-bound and Zn-free His residues. Using several model peptides and proteins, we find that all Zn-bound His residues have substantially lower HDX reaction rates in the presence of the metal. The vast majority of non-Zn-binding His residues undergo no significant changes in HDX reaction rates when their reactivity is compared in the presence and absence of Zn. Using this new approach, we then determined the Zn binding site of β-2-microglobulin, a protein associated with metal-induced amyloidosis. Together, these results suggest that HDX-MS of His C2-hydrogens is a promising new method for identifying Zn-bound histidines in metalloproteins.

• Authors: Jia Dong, Katie L Callahan, Nicholas B Borotto, Richard W Vachet
• Journal: Analytical chemistry (Anal Chem) Vol. 86 Issue 1 Pg. 766-73 (Jan 07 2014) ISSN: 1520-6882 [Electronic] United States
• PMID: 24313328 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Metalloproteins
• Histidine
• Zinc

Topics

• Amino Acid Sequence
• Animals
• Binding Sites (physiology)
• Cattle
• Crystallography, X-Ray
• Deuterium Exchange Measurement (methods)
• Histidine (analysis, genetics, metabolism)
• Humans
• Mass Spectrometry (methods)
• Metalloproteins (analysis, genetics, metabolism)
• Molecular Sequence Data
• Protein Structure, Secondary
• Zinc (analysis, metabolism)