Purification of the 22 kDa protein substrate of botulinum ADP‐ribosyltransferase C3 from porcine brain cytosol and its characterization as a GTP‐binding protein highly homologous to the rho gene product
- 16 January 1989
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 243 (1), 70-76
- https://doi.org/10.1016/0014-5793(89)81220-7
Abstract
The 22 kDa protein substrate of botulinum ADP-ribosyltransferase C3 was purified from porcine brain cytosol by acetone precipitation, CM-Sephadex, octyl-Sepharose and TSK phenyl-5PW HPLC chromatography to apparent homogeneity. ADP-ribosylation of the protein was increased by guanine nucleotides (GTP, GDP, GTPγS, each 100μM) but not by GMP, ATP or ATPγS. The purified 22 kDa protein bound maximally 0.9 mol [35S]GTPγS and hydrolyzed GTP with the rate 0.007 mol per mol protein. Amino acid sequences were obtained from two tryptic peptides, selected from an in situ digestion of Immobilon electrotransferred, gel purified ADP-ribosylated substrate. The two sequences obtained, cover 23 residues from the corresponding sequences in human rho.Keywords
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