Purification and structural characterization of the putative gag-pol protease of human immunodeficiency virus.
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.
Lillehoj
E P
EP
Laboratory of Molecular Microbiology, Program Resources, Inc., Frederick Cancer Research Facility, Maryland 21701.
Salazar
F H
FH
Mervis
R J
RJ
Raum
M G
MG
Chan
H W
HW
Ahmad
N
N
Venkatesan
S
S
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000302:isolation & purification*; D015683:Gene Products, gag; D005814:Genes, Viral; D006678:HIV / Q000201:enzymology*; D016333:HIV Protease; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012191:Retroviridae Proteins / Q000302:isolation & purification
D015683:Gene Products, gag; D012191:Retroviridae Proteins; D010450:Endopeptidases; D016333:HIV Protease