Formation of polyamine-modified peptides during protein digestion.
The effect of polyamines on the digestion of proteins by serine proteases was examined. Based on the mechanism of action of serine proteases, it was anticipated that nucleophilic functionalities such as amino groups in polyamine, rather than hydroxyl ions, would react with peptide bonds during the hydrolysis process. If this were the case, polyamine might be covalently linked to the C-terminus of the product peptides during protein digestion. In order to test this hypothesis, hemoglobin was used as a model protein and was digested with trypsin in the presence of polyamine. The product peptides were separated, collected by HPLC, and analyzed by MALDI-TOF MS using post-source decay. The results showed that some peptides were indeed modified with polyamine at the C-terminus.
Ito
Toshiyuki
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama 350 0295, Japan.
Sugita
Yoshiaki
Y
Ikeguchi
Yoshihiko
Y
Shirahata
Akira
A
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D008956:Models, Chemical; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry*; D011073:Polyamines / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D006454:Hemoglobins; D010446:Peptide Fragments; D011073:Polyamines; D014357:Trypsin