Occurrence of an HIV-1 gp160 endoproteolytic activity in low-density vesicles and evidence for a distinct density distribution from endogenously expressed furin and PC7/LPC convertases.
Human immunodeficiency virus (HIV) glycoprotein (gp) 160 processing by host cell proteinases is an essential step for viral fusion and infectivity. We have identified a rat liver subcellular fraction which specifically processes gp160 into gp120 and gp41. Using equilibration of microsomes in sucrose gradients, the gp160 cleavage activity was associated with particles equilibrating at low densities, well-separated from the endoplasmic reticulum, cis-Golgi network, Golgi stacks, lysosomes and plasma membrane. Its density distribution was compatible with light secretory vesicles derived from the trans-Golgi network (TGN) or to endosomes, but association with endosomes was not supported by free flow electrophoresis. Although furin and pro-protein convertase (PC) 7/LPC have been proposed as the major gp160 processing convertases, the rat liver microsomal gp160 processing activity was essentially resolved from furin and only partially overlapped PC7/LPC. These data suggest that proteinase(s) other than furin and PC7/LPC, presumably located in TGN-derived vesicles, may participate in the gp160 processing into gp120 and gp41.
Wouters
S
S
Laboratoire de Chimie Physique des Macromolécules aux Interfaces, Université Libre de Bruxelles, Brussels, Belgium. swouters@ulb.ac
Decroly
E
E
Vandenbranden
M
M
Shober
D
D
Fuchs
R
R
Morel
V
V
Leruth
M
M
Seidah
N G
NG
Courtoy
P J
PJ
Ruysschaert
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D002451:Cell Compartmentation; D002499:Centrifugation, Density Gradient; D004586:Electrophoresis / Q000379:methods; D011992:Endosomes / Q000378:metabolism; D045683:Furin; D006056:Golgi Apparatus / Q000378:metabolism; D019302:HIV Envelope Protein gp160 / Q000737:chemistry / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry*; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D017421:Sequence Analysis; D012697:Serine Endopeptidases / Q000378:metabolism*; D013347:Subcellular Fractions; D013381:Subtilisins / Q000378:metabolism*
D019302:HIV Envelope Protein gp160; D010446:Peptide Fragments; C490931:Pcsk7 protein, rat; D012697:Serine Endopeptidases; D013381:Subtilisins; C107583:proprotein convertase PC7; D045683:Furin