Characterization of succinimide-type dehydration products of recombinant hirudin variant 1 by electrospray tandem mass spectrometry.
Two congeners Q4 and Q5 inferred in earlier analyses to be cyclic succinimide-type dehydration products of recombinant hirudin (variant 1) were structurally fully characterized. After isotopic labeling by ring-opening with H2(18)O, the suspected anhydro-positions Asp53 and Asp33 were confirmed by tandem mass spectrometry (MS/MS) sequencing of relevant smaller peptides directly in the enzymatic hydrolysates (V8 protease) using electrospray MS/MS. The chosen strategy proved highly efficient and sensitive.
Grossenbacher
H
H
Pharmaceutical Division, Ciba-Geigy AG, Basel, Switzerland.
Märki
W
W
Coulot
M
M
Müller
D
D
Richter
W J
WJ
D006629:Hirudins / Q000096:biosynthesis*; D013058:Mass Spectrometry / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae; D013388:Succinimides
D006629:Hirudins; D011994:Recombinant Proteins; D013388:Succinimides; C072298:hirudin HV1