E. Canovadavis et al., CONFIRMATION BY MASS-SPECTROMETRY OF A TRISULFIDE VARIANT IN METHIONYL HUMAN GROWTH-HORMONE BIOSYNTHESIZED IN ESCHERICHIA-COLI, Analytical chemistry, 68(22), 1996, pp. 4044-4051
A sulfur-containing compound found in acid hydrolysates of proteins wa
s identified 30 years ago as a trisulfide: bis-(2-amino-2-carboxyethyl
) trisulfide (cysteine(2)S(3)). At that time, studies concerning the c
hemistry of sulfur-transferring enzyme systems suggested that cysteine
(2)S(3) also existed in biological systems, Two decades later, a cysti
ne trisulfide structure was postulated in the regulator protein molecu
le for the activation of delta-aminolevulinate synthetase, Recently, a
trisulfide bond was reported to occur in the minor loop disulfide at
Cys(182)-Cys(189) in human growth hormone, Wet have detected a trisulf
ide structure in methionyl human growth hormone iu the major loop disu
lfide Cys(53)-Cys(165). The development of mass spectral analyses of h
igh molecular weight molecules, such as proteins, led to the eventual
identification of the modification, A tandem mass spectral analysis on
a Sciex electrospray instrument localized an addition of 32 Da to the
Cys(53)-Cys(165) fragment, Elemental composition was determined by ac
curate mass measurement obtained by peak matching to a synthetic pepti
de and established that an extra sulfur atom was involved.