Purification and characterization of the external envelope glycoprotein from two human immunodeficiency virus type 1 variants, HTLV-IIIB and HTLV-IIIRF.
External envelope glycoprotein from cell membranes and culture media of H9 cells infected with human immunodeficiency virus type 1 (HIV-1) isolate HTLV-IIIRF was isolated by immunoaffinity chromatography and compared with similar materials isolated from another variant, HTLV-IIIB. Envelope glycoprotein from IIIB and IIIRF appears to be identical, whether isolated from infected cell membranes or culture media. The molecular size of the IIIRF external envelope glycoprotein was 110 kilodaltons, whereas the relative size of IIIB gp120 was 123 kilodaltons. Amino-terminal sequence analysis of purified external envelope glycoprotein isolated from infected cell membranes or culture fluids revealed identical single sequences for the first 20 amino acids for each variant. The sequences obtained for IIIB gp120 were identical to those reported for the BH10 clone of the IIIB isolate, and the sequences determined for IIIRF gp110 matched the amino acid sequence predicted for the HAT3 clone of the Haitian HIV isolate. The amino-terminal sequences of external envelope glycoproteins isolated from either HIV-1 variant corresponded to the sequence starting at the proposed proteolytic cleavage site for the processing of the signal peptide of gp160. Immunization with external envelope glycoprotein isolated from either of the two HIV-1 variants yielded goat antibodies that primarily precipitated the homologous antigen. Sequential immunization of a single goat with gp120 and then gp110 resulted in the generation of antibodies that precipitated external envelope glycoprotein from both variants.
Pyle
S W
SW
Program Resources, Inc., Frederick, Maryland 21701.
Dubois
G C
GC
Robey
W G
WG
Bess
J W
JW
Fischinger
P J
PJ
Arthur
L O
LO
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology; D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D003470:Culture Media / Q000032:analysis; D006678:HIV / Q000032:analysis* / Q000145:classification / Q000276:immunology; D015483:HIV Antibodies; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000276:immunology / Q000302:isolation & purification*
D000914:Antibodies, Viral; D003470:Culture Media; D015483:HIV Antibodies; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins