Isolation and characterization of porcine somatotropin containing a succinimide residue in place of aspartate129.
Aspartate129 in porcine somatotropin was converted into a cyclic imide residue (succinimide) under acidic solution conditions. Reversed-phase high performance liquid chromatography was utilized to isolate and quantitate this altered species, which accounted for approximately 30% of the total protein. The molecular mass of this modified species was determined by electrospray mass spectrometry to be 18 Da less than normal porcine somatotropin, indicative of a loss of 1 H2O molecule. Tryptic peptide mapping demonstrated that the peptide composed of residues 126-133 was altered in this modified protein. Amino acid analysis, amino acid sequencing, mass spectrometry, and capillary zone electrophoresis were used to demonstrate that aspartate129 in this peptide had been converted into a succinimide residue. Further confirmation that this peptide contained a succinimide was obtained by hydrolyzing the modified peptide at pH 9.0, which yielded both the aspartate and isoaspartate peptides.
Violand
B N
BN
Animal Sciences Division, Monsanto Company, St. Louis, Missouri 63198.
Schlittler
M R
MR
Kolodziej
E W
EW
Toren
P C
PC
Cabonce
M A
MA
Siegel
N R
NR
Duffin
K L
KL
Zobel
J F
JF
Smith
C E
CE
Tou
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D013004:Somatostatin / Q000737:chemistry* / Q000302:isolation & purification; D013388:Succinimides / Q000032:analysis*; D013552:Swine; D014357:Trypsin
D010446:Peptide Fragments; D013388:Succinimides; C032620:succinimide; D001224:Aspartic Acid; D013004:Somatostatin; D014357:Trypsin