title: Peptide bond cleavage site determination of novel proteolytic enzymes found in ROS 17/2.8 cell lysates.
abstract: We have identified proteolytic activities in the rat osteoblastic osteosarcoma cell line ROS 17/2.8 which are capable of cleaving a peptide substrate for protein kinase C-mediated phosphorylation (PSPKC, Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys). Using polyacrylamide gel electrophoresis conditions similar to those used to resolve small molecular weight proteins, the peptide bonds of PSPKC which are cleaved by the proteolytic activities present in ROS 17/2.8 cell lysates have been determined. These activities cleave the Ser-Arg, Thr-Leu, and Ser-Val peptide bonds. To date, no proteolytic activities present in osteoblast cell lysates have been described with the aforementioned peptide bond specificities, suggesting that these activities are novel. The PSPKC-cleaved peptide fragment pattern generated was similar for several different osteoblast cell lysates. Lysates generated from different rat tissues were also able to cleave PSPKC, but the peptide fragment pattern generated by ROS 17/2.8 cell lysates appeared to be unique amongst these tissues.
authors.affiliation: Department of Biology, Seton Hall University, South Orange, New Jersey 07079, USA.
authors.forename: P T
authors.initials: PT
authors.lastname: Guidon
authors.forename: D
authors.initials: D
authors.lastname: Perrin
authors.forename: P
authors.initials: P
authors.lastname: Harrison
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003470:Culture Media; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D010006:Osteoblasts / Q000201:enzymology*; D010446:Peptide Fragments / Q000378:metabolism; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
chemical_list: D003470:Culture Media; D009842:Oligopeptides; D010446:Peptide Fragments; D011493:Protein Kinase C; D010447:Peptide Hydrolases