Expression and purification of glutathione S-transferase-tagged HIV-1 gp120: no evidence of an interaction with CD26.
We describe the use of a new baculovirus expression vector to enable the secretion of the major surface glycoprotein of HIV-1 (gp120) fused to the carboxy-terminus of the widely used affinity tag glutathione S-transferase. The secreted protein can be purified in a single step with the minimum of denaturation on immobilised glutathione and is as active as the parental molecule in binding CD4. We use this molecule in a variety of assay formats to examine the gp120 interaction with CD26, a reported auxiliary molecule in the HIV entry process. We find no evidence of a CD26-gp120 interaction in the absence or presence of CD4.
Wang
Y H
YH
NERC Institute of Virology, Oxford, United Kingdom.
Davies
A H
AH
Jones
I M
IM
D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D015699:HIV Envelope Protein gp120; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D018819:Dipeptidyl Peptidase 4