The use of plasma desorption time-of-flight mass spectrometry to screen for products of prohormone processing in crude tissue extracts.
Californium-252 plasma desorption mass spectrometry (252Cf PDMS) of a crude, desalted, extract of piscine endocrine pancreas provided mass information for the major biologically active peptide hormones present in this tissue. An extraction procedure compatible with 252Cf PDMS analysis was developed. In extracts of catfish pancreas, strong molecular ions were identified in the positive mode for somatostatin-14 (1638 amu), O-glycosylated somatostatin-22 (2944 amu), glucagon (3512 amu), glucagon-like peptide (3785 amu), insulin (ca. 5550 amu), and other prohormone-derived peptides. Both protonated species and sodium adducts were apparent in the mass spectrum. A number of other molecular ions were observed including somatostatin-26, 1-10 (1014 amu) and the entire portion of prosomatostatin-22 remaining after removal of somatostatin-22 (6465 amu). The data obtained by this method also resulted in the identification of the third major product of proglucagon processing in catfish pancreas, glicentin-related polypeptide. Subtractive Edman degradation analyzed by 252Cf PDMS was also used to confirm a mass assignment.
Andrews
P C
PC
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.
Alai
M
M
Cotter
R J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D002142:Californium; D002397:Catfishes; D007515:Islets of Langerhans / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000302:isolation & purification*; D011498:Protein Precursors / Q000302:isolation & purification*
D010455:Peptides; D011498:Protein Precursors; D002142:Californium