4th International AIDS Conference Stockholm, Sweden. — June 12-16, 1988

Int Conf AIDS. 1988 Jun 12-16;4:1.114 (abstract no. 1008)

Bharat Parekh and Roger Walker
Bio-Rad Laboratories, Clinical Division, 1000 Alfred Nobel Drive, Hercules, CA 94547, USA

OBJECTIVE: To isolate and characterize HIV-1 glycoproteins.

METHODS: HIV-1 glycoproteins were separated from nonglycosylated components from detergent-disrupted virus using a lentil lectin column. Pools were analyzed on a Western Blot. Non-reducing gels were run to examine inter- and intramolecular disulfide linkages in these glycoproteins.

RESULTS: On a lentil lectin column, non-glycosylated proteins eluted in the void volume peak. Bound glycoproteins were eluted with a buffer containing a-methyl mannoside and a-methyl glucoside. Analysis of these two peaks by Western Blot, using a polyclonal human serum, revealed that Peak 1 contained all of the non-glycosylated components, while Peak 2 had most of the viral glycoproteins. Non-reducing SDS-PAGE revealed that gp120 and gp41-43 are not disulfide-linked. However, an additional immunoreactive band was seen at about 90Kd which may indicate that a proportion of gp41-43 may exist as a dimer in the virion. Non-reduced gp41-43 was more immunoreactive than the reduced molecule, suggesting the importance of intramolecular disulfide bonds in maintaining the folding and conformation of antigenic sites on this glycoprotein.

CONCLUSION: Although gp120 and gp41-43 are not disulfide-linked, a small proportion of gp41-43 may exist as a dimer. The non-reduced form of gp41-43 was found to be more immunoreactive than the reduced molecule.

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