Expression of recombinant pro-neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by 'prohormone thiol protease' (PTP).

Authors

Martin R. Schiller, University of Nevada, Las VegasFollow
Andrea B. Kohn, Uniformed Services University of the Health Sciences
Liane M. Mende-Mueller, Medical College of Wisconsin
Kurt Miller, University of WyomingFollow
Vivian Y. Hook, University of California - San DiegoFollow

Document Type

Article

Publication Date

3-11-1996

Publication Title

FEBS Letters

Volume

382

Issue

1-2

First page number:

6

Last page number:

10

Abstract

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), proopiomelanocortin (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.

Keywords

Amino acid sequence; Animals; Base Sequence; Biosynthesis; Cysteine Endopeptidases/metabolism; Endopeptidases; Enkephalins; Enkephalins/biosynthesis; Enkephalins/genetics; Enkephalins/isolation & purification; Enkephalins/metabolism; Escherichia coli; Escherichia coli/genetics; Genetics; Molecular Sequence Data; Nucleotide sequence; Neuropeptide Y; Neuropeptide Y/biosynthesis; Neuropeptide Y/genetics; Neuropeptide Y/isolation & purification; Neuropeptide Y/metabolism; Proopiomelanocortin; Pro-Opiomelanocortin/biosynthesis; Pro-Opiomelanocortin/genetics; Pro-Opiomelanocortin/isolation & purification; Pro-Opiomelanocortin/metabolism; Proprotein convertases; Protein precursors; Protein Precursors/biosynthesis; Protein Precursors/genetics; Protein Precursors/isolation & purification; Protein Precursors/metabolism; Protein Processing; Post-Translational; Proteins—Synthesis; Rats; Recombinant Fusion Proteins/biosynthesis; Recombinant Fusion Proteins/isolation & purification; Recombinant Fusion Proteins/metabolism; Substrate Specificity; Swine

Disciplines

Biochemistry | Life Sciences | Medicine and Health Sciences | Molecular Biology

Language

English

Repository Citation

Schiller, M. R., Kohn, A. B., Mende-Mueller, L. M., Miller, K., Hook, V. Y. (1996). Expression of recombinant pro-neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by 'prohormone thiol protease' (PTP).. FEBS Letters, 382(1-2), 6-10.
http://dx.doi.org/10.1016/0014-5793(96)00083-X