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Peptide bond cleavage site determination of novel proteolytic enzymes found in ROS 17/2.8 cell lysates

✍ Scribed by Peter T. Guidon Jr.; Dana Perrin; Patricia Harrison


Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
424 KB
Volume
166
Category
Article
ISSN
0021-9541

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✦ Synopsis


South Orange, New )ersey 07079

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 17/23 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, sug- gesting 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. o 1996 WiIey-Liss, Inc.