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Cleavage of recombinant and cell derived human immunodeficiency virus 1 (HIV‐1) Nef protein by HIV‐1 protease
Author(s) -
Gaedigk-Nitschko K.,
Schön A.,
Wachinger G.,
Erfle V.,
Kohleisen B.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)01370-g
Subject(s) - protease , cleavage (geology) , recombinant dna , ns2 3 protease , hiv 1 protease , biology , monoclonal antibody , virology , microbiology and biotechnology , biochemistry , chemistry , antibody , enzyme , gene , paleontology , fracture (geology) , immunology
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.