Cloning, bacterial expression, and characterization of the Mason-Pfizer monkey virus proteinase

O. Hruskova-Heidingsfeldova, Martin Andreansky, M. Fabry, I. Blaha, P. Strop, E. Hunter

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

We have cloned and expressed the 3' region of the Mason-Pfizer monkey virus pro gene in Escherichia coli. The recombinant 26-kDa precursor undergoes rapid self-processing both in E. coli and in vitro at the NH2 terminus, yielding a proteolytically active 17-kDa protein, p17. This initial cleavage is followed in vitro by a much slower self-processing that leads to emergence of proteolytically active p12 and a COOH-terminal cleavage product p5. We have found the NH2-terminal processing site of both the p17 and p12 to be identical and similar to the amino terminus of the mouse mammary tumor virus proteinase. We have also identified the COOH-terminal processing site of the p12 form. Using purified recombinant proteins and synthetic oligopeptide substrates based on naturally occurring retroviral processing sites, we have determined the enzymatic activity and specificity of the Mason-Pfizer monkey virus proteinase to be more closely related to that of myeloblastosis-associated virus proteinase rather than that of the Human immunodeficiency virus type 1 proteinase. Inhibition studies using peptide inhibitors support these results.

Original languageEnglish
Pages (from-to)15053-15058
Number of pages6
JournalJournal of Biological Chemistry
Volume270
Issue number25
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Mason-Pfizer monkey virus
Cloning
Viruses
Organism Cloning
Peptide Hydrolases
Processing
Escherichia coli
Mouse mammary tumor virus
Satellite Viruses
Oligopeptides
Recombinant Proteins
HIV-1
Tumors
Peptides
Genes
Substrates
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cloning, bacterial expression, and characterization of the Mason-Pfizer monkey virus proteinase. / Hruskova-Heidingsfeldova, O.; Andreansky, Martin; Fabry, M.; Blaha, I.; Strop, P.; Hunter, E.

In: Journal of Biological Chemistry, Vol. 270, No. 25, 01.01.1995, p. 15053-15058.

Research output: Contribution to journalArticle

Hruskova-Heidingsfeldova, O. ; Andreansky, Martin ; Fabry, M. ; Blaha, I. ; Strop, P. ; Hunter, E. / Cloning, bacterial expression, and characterization of the Mason-Pfizer monkey virus proteinase. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 25. pp. 15053-15058.
@article{3f115b9e8ac447b0a2b9d70351347e23,
title = "Cloning, bacterial expression, and characterization of the Mason-Pfizer monkey virus proteinase",
abstract = "We have cloned and expressed the 3' region of the Mason-Pfizer monkey virus pro gene in Escherichia coli. The recombinant 26-kDa precursor undergoes rapid self-processing both in E. coli and in vitro at the NH2 terminus, yielding a proteolytically active 17-kDa protein, p17. This initial cleavage is followed in vitro by a much slower self-processing that leads to emergence of proteolytically active p12 and a COOH-terminal cleavage product p5. We have found the NH2-terminal processing site of both the p17 and p12 to be identical and similar to the amino terminus of the mouse mammary tumor virus proteinase. We have also identified the COOH-terminal processing site of the p12 form. Using purified recombinant proteins and synthetic oligopeptide substrates based on naturally occurring retroviral processing sites, we have determined the enzymatic activity and specificity of the Mason-Pfizer monkey virus proteinase to be more closely related to that of myeloblastosis-associated virus proteinase rather than that of the Human immunodeficiency virus type 1 proteinase. Inhibition studies using peptide inhibitors support these results.",
author = "O. Hruskova-Heidingsfeldova and Martin Andreansky and M. Fabry and I. Blaha and P. Strop and E. Hunter",
year = "1995",
month = "1",
day = "1",
doi = "10.1074/jbc.270.25.15053",
language = "English",
volume = "270",
pages = "15053--15058",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "25",

}

TY - JOUR

T1 - Cloning, bacterial expression, and characterization of the Mason-Pfizer monkey virus proteinase

AU - Hruskova-Heidingsfeldova, O.

AU - Andreansky, Martin

AU - Fabry, M.

AU - Blaha, I.

AU - Strop, P.

AU - Hunter, E.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - We have cloned and expressed the 3' region of the Mason-Pfizer monkey virus pro gene in Escherichia coli. The recombinant 26-kDa precursor undergoes rapid self-processing both in E. coli and in vitro at the NH2 terminus, yielding a proteolytically active 17-kDa protein, p17. This initial cleavage is followed in vitro by a much slower self-processing that leads to emergence of proteolytically active p12 and a COOH-terminal cleavage product p5. We have found the NH2-terminal processing site of both the p17 and p12 to be identical and similar to the amino terminus of the mouse mammary tumor virus proteinase. We have also identified the COOH-terminal processing site of the p12 form. Using purified recombinant proteins and synthetic oligopeptide substrates based on naturally occurring retroviral processing sites, we have determined the enzymatic activity and specificity of the Mason-Pfizer monkey virus proteinase to be more closely related to that of myeloblastosis-associated virus proteinase rather than that of the Human immunodeficiency virus type 1 proteinase. Inhibition studies using peptide inhibitors support these results.

AB - We have cloned and expressed the 3' region of the Mason-Pfizer monkey virus pro gene in Escherichia coli. The recombinant 26-kDa precursor undergoes rapid self-processing both in E. coli and in vitro at the NH2 terminus, yielding a proteolytically active 17-kDa protein, p17. This initial cleavage is followed in vitro by a much slower self-processing that leads to emergence of proteolytically active p12 and a COOH-terminal cleavage product p5. We have found the NH2-terminal processing site of both the p17 and p12 to be identical and similar to the amino terminus of the mouse mammary tumor virus proteinase. We have also identified the COOH-terminal processing site of the p12 form. Using purified recombinant proteins and synthetic oligopeptide substrates based on naturally occurring retroviral processing sites, we have determined the enzymatic activity and specificity of the Mason-Pfizer monkey virus proteinase to be more closely related to that of myeloblastosis-associated virus proteinase rather than that of the Human immunodeficiency virus type 1 proteinase. Inhibition studies using peptide inhibitors support these results.

UR - http://www.scopus.com/inward/record.url?scp=0029043595&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029043595&partnerID=8YFLogxK

U2 - 10.1074/jbc.270.25.15053

DO - 10.1074/jbc.270.25.15053

M3 - Article

VL - 270

SP - 15053

EP - 15058

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 25

ER -