Expression of recombinant pro‐neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by ‘prohormone thiol protease’ (PTP)

FEBS Letters

Miller, Kurt¹  Schiller, Martin R²  Kohn, Andrea B²  Hook, Vivian Y.H³  Mende-Mueller, Liane M⁴ 

[1] Department of Molecular Biology, University of Wyoming, Laramie, WY, USA;Department of Biochemistry, Uniformed Services University, Bethesda, MD, USA;Department of Medicine, University of California, San Diego, CA, USA;Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA

关键词: Recombinant prohormone;    Proprotein processing;    Cysteine protease;    PTP;    prohormone thiol protease;    Pro-NPY;    proneuropeptide Y;    POMC;    proopiomelanocortin;    PE;    proenkephalin;    PC;    prohormone convertase;    ACTH;    adrenocorticotropin hormone;    IPTG;    isopropyl-ß-thiogalactopyranoside;   

DOI  :  10.1016/0014-5793(96)00083-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The preference of the ‘prohormone thiol protease’ (PTP), a candidate prohormone processing enzyme, for different peptide precursors was assessed in vitro with recombinant prohormones near estimated in vivo levels. Pro-neuropeptide Y (pro-NPY), prooplomelanocortin (POMC), and proenkephalin (PE) were expressed at high levels in E. coli. Purification of prohormones utilized a combination of DEAE-Sepharose, Mono Q, and preparative electrophoresis. PTP cleaved PE most readily, and also cleaved pro-NPY. The processing of POMC by PTP was minimal. These results demonstrate PTP's preference for certain prohormone substrates.

【 授权许可】

Unknown