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. 1991 Aug 11;19(15):4253–4257. doi: 10.1093/nar/19.15.4253

Characterization of the CRPCY core formed after treatment with carboxypeptidase Y.

Z H Yang 1, S Bobin 1, J S Krakow 1
PMCID: PMC328570  PMID: 1651482

Abstract

CRP is resistant to attack by carboxypeptidase Y at 37 degrees C, whereas cAMP-CRP is digested yielding a core terminating at Thr-202 and lacking the seven carboxyl-terminal amino acid residues. A similar core (CRPCY) is formed when CRP is incubated with carboxypeptidase Y at 47 degrees C in the absence of cAMP. CRPCY has a more open conformation than CRP at 37 degrees C. While unliganded CRP is resistant to trypsin, CRPCY is sensitive to tryptic attack. Dithionitrobenzoic acid-mediated intersubunit disulfide crosslinking of CRP requires cAMP, CRPCY subunits are crosslinked in the absence of cAMP. The carboxyl-terminal region of unliganded CRP is conformationally restricted at 37 degrees C. The CRPCY retains cAMP binding activity. The CRPCY which terminates at Thr-202, no longer binds lac P+ DNA nor stimulates abortive initiation by RNA polymerase from the lac P+ promoter. The results indicate that the C-terminal region of CRP participates in the conformational stability of the closed form of CRP and indirectly in DNA binding by the open cAMP-CRP conformer.

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