Folding of HIV-1 envelope glycoprotein involves extensive isomerization of disulfide bonds and conformation-dependent leader peptide cleavage.
Human immunodeficiency virus binds and enters cells via the Envelope glycoprotein gp160 at its surface. In infected cells, gp160 is found not only on the plasma membrane but also in the endoplasmic reticulum (ER). Our aim was to establish rate-determining steps in the maturation process of gp160, using a radioactive pulse-chase approach. We found that gp160 has an intricate folding process: disulfide bonds start to form during synthesis but undergo extensive isomerization until the correct native conformation is reached. Removal of the leader peptide critically depends on formation of at least some disulfide bonds in subunit gp120 during folding. Envelope folds extremely slowly and therefore resides in the ER longer than other proteins, but the yield of properly folded molecules is high and degradation is undetectable. The large quantity of gp160 in the ER hence is a result of its slow transit through this compartment. We show here that newly synthesized HIV-1 Envelope glycoprotein apparently follows a slow but high-yield folding path in which co- and post-translational formation of disulfide bonds in gp120, disulfide isomerization and conformation dependent removal of the leader sequence are determining and intertwined events.
Land
Aafke
A
Department Bio-Organic Chemistry 1, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Zonneveld
Duco
D
Braakman
Ineke
I
D004220:Disulfides / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D019302:HIV Envelope Protein gp160 / Q000737:chemistry* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007536:Isomerism; D007700:Kinetics; D011487:Protein Conformation; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism*; D021382:Protein Sorting Signals
D004220:Disulfides; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D021382:Protein Sorting Signals