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Structural characterization of human hemoglobin crosslinked by bis(3,5‐dibromosalicyl) fumarate using mass spectrometric techniques
Author(s) -
Yu Zhonghua,
Friso Giulia,
Miranda Jj,
Burlingame Alma L.,
Patel Mehul J.,
LoTseng Thomas,
Moore Edwin G.
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560061209
Subject(s) - characterization (materials science) , hemoglobin , chemistry , mass spectrometry , chromatography , combinatorial chemistry , materials science , nanotechnology , biochemistry
Diaspirin crosslinked hemoglobin (DCLHb) was analyzed by mass spectrometric‐based techniques to identify the protein modifications effected by the crosslinking reaction with bis(3,5‐dibromosalicyl) fumarate. DCLHb consists of two principal components. These components were isolated by size‐exclusion chromatography and identified by measurement of their molecular weight using electrospray mass spectrometry and subsequent peptide mass mapping and mass spectrometric sequence analysis of their individual digests. Three major RP‐HPLC fractions were observed from the major hemoglobin in DCLHb. Their MWs matched the MW of heme, intact hemoglobin β‐chain, and two hemoglobin α‐chains crosslinked by a fumarate moiety, respectively. The minor HPLC peaks of DCLHb were also separated, and characterized by mass spectrometric methods. These minor components revealed additional details of the structural nature of covalent modification of DCLHb.