Quantitative analysis of pyroglutamic acid in peptides.
A simplified and rapid procedure for the determination of pyroglutamic acid in peptides was developed. The method involves the enzymatic cleavage of an N-terminal pyroglutamate residue using a thermostable pyroglutamate aminopeptidase and isocratic HPLC separation of the resulting enzymatic hydrolysate using a column switching technique. Pyroglutamate aminopeptidase from a thermophilic archaebacteria, Pyrococcus furiosus, cleaves N-terminal pyroglutamic acid residue independent of the molecular weight of the substrate. It cleaves more than 85% of pyroglutamate from peptides whose molecular weight ranges from 362.4 to 4599.4 Da. Thus, a new method is presented that quantitatively estimates N-terminal pyroglutamic acid residue in peptides.
Suzuki
Y
Y
Nisshin Flour Milling Company, Limited, Saitama, Japan. suzukiyo@mail.ni-net.co.jp
Motoi
H
H
Sato
K
K
D029845:Amphibian Proteins; D001407:Bacillus / Q000201:enzymology; D001839:Bombesin / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005755:Gastrins / Q000737:chemistry; D007987:Gonadotropin-Releasing Hormone / Q000737:chemistry; D006728:Hormones / Q000737:chemistry*; D006801:Humans; D036361:Peptide Hormones; D010455:Peptides / Q000737:chemistry*; D019711:Pyrococcus / Q000201:enzymology; D011751:Pyroglutamyl-Peptidase I; D011761:Pyrrolidonecarboxylic Acid / Q000032:analysis*; D013373:Substance P / Q000737:chemistry; D013973:Thyrotropin-Releasing Hormone / Q000737:chemistry
D029845:Amphibian Proteins; D005755:Gastrins; D006728:Hormones; D036361:Peptide Hormones; D010455:Peptides; D013373:Substance P; D007987:Gonadotropin-Releasing Hormone; D013973:Thyrotropin-Releasing Hormone; C025727:sauvagine; C036227:gastrin I; D011751:Pyroglutamyl-Peptidase I; D001839:Bombesin; D011761:Pyrrolidonecarboxylic Acid