Purification and structural characterization of the putative gag-pol protease of human immunodeficiency virus

E. P. Lillehoj, F. H.R. Salazar, R. J. Mervis, M. G. Raum, H. W. Chan, N. Ahmad, S. Venkatesan

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

We have purified a 10,774-dalton protein from human immunodeficiency virus (HIV) type 1 that is encoded in the protease domain of the pol open reading frame (ORF). Radiochemical amino acid microsequencing identified 12 amino acids from the stretch of 39 N-terminal residues of this protein, beginning with a PQITLW sequence at position 69 of the pol ORF. Radiosequencing of selected tryptic peptides of the protein identified 11 additional residues (Leu-9 and Val-2) in six peptides encompassing the entire molecule of 99 residues. A protein of similar size and identical N-terminal sequence (determined through the first 39 residues) was present among the processed HIV pol gene products in Escherichia coli which expressed the entire HIV pol ORF. The C terminus of both the viral and E. coli-expressed proteins was inferred to be contiguous with the N terminus of the p64-p51 reverse transcriptase on the basis of tryptic mapping and specific immunoreactivity with an antiserum against a dodecapeptide located upstream of the reverse transcriptase. Thus, the initial processing of the pol precursor that generates the native protease is apparently preserved across phylogenetic barriers. Although the purified viral protease lacked measurable proteolytic activity, the bacterial extracts were capable of processing an HIV gag precursor protein synthesized in E. coli.

Original languageEnglish (US)
Pages (from-to)3053-3058
Number of pages6
JournalJournal of virology
Volume62
Issue number8
StatePublished - Jan 1 1988
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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    Lillehoj, E. P., Salazar, F. H. R., Mervis, R. J., Raum, M. G., Chan, H. W., Ahmad, N., & Venkatesan, S. (1988). Purification and structural characterization of the putative gag-pol protease of human immunodeficiency virus. Journal of virology, 62(8), 3053-3058.