Cleavage of Recombinant and Cell Derived Human Immunodeficiency Virus 1 (HIV-1) Nef Protein by HIV-1 Protease

FEBS Lett. 1995 Jan 9;357(3):275-8. doi: 10.1016/0014-5793(94)01370-g.

Abstract

Recombinant purified Nef protein of HIV-1, as well as Nef protein derived from extracts of permanently HIV-1 infected glioblastoma cells and monocytes, are specifically cleaved by the HIV-1 protease. Nef cleavage products in cellular extracts treated with protease showed identical molecular weights as those obtained by digestion of purified Nef with recombinant HIV-1 protease. Since cellular extracts were prepared by detergent and mechanical lysis it cannot be excluded that physiological cytoplasmic conditions were altered. The lack of Nef cleavage by endogenous HIV-1 protease in infected cells might be due to low concentrations of viral protease and the presence of Gag precursor molecules as natural substrate. Using a panel of monoclonal antibodies two cleavage fragments of 19 kDa and 8 kDa were defined. The cleavage site was located by microsequencing between amino acid 57 and 58 (AW*LEAQEEEEVGF). The conserved cleavage motif within HIV-1 Nef suggests a potential biological function of Nef processing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal
  • Gene Products, gag / metabolism
  • Gene Products, nef / metabolism*
  • HIV Protease / metabolism*
  • HIV-1 / enzymology
  • HIV-1 / metabolism*
  • Hydrolysis
  • Molecular Sequence Data
  • Tumor Cells, Cultured
  • nef Gene Products, Human Immunodeficiency Virus

Substances

  • Antibodies, Monoclonal
  • Gene Products, gag
  • Gene Products, nef
  • nef Gene Products, Human Immunodeficiency Virus
  • HIV Protease