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J Virol, August 1998, p. 6537-6545, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Moloney Murine Leukemia Virus Envelope Protein
Subunits, gp70 and Pr15E, Form a Stable Disulfide-Linked
Complex
Dirk-Jan E.
Opstelten,*
Michael
Wallin, and
Henrik
Garoff
Department for Biosciences at Novum,
Karolinska Institute, S-141 57 Huddinge, Sweden
Received 29 January 1998/Accepted 30 April 1998
The nature and stability of the interactions between the gp70 and
Pr15E/p15E molecules of murine leukemia virus (MLV) have been disputed
extensively. To resolve this controversy, we have performed
quantitative biochemical analyses on gp70-Pr15E complexes formed after
independent expression of the amphotropic and ecotropic Moloney MLV
env genes in BHK-21 cells. We found that all
cell-associated gp70 molecules are disulfide linked to Pr15E whereas
only a small amount of free gp70 is released by the cells. The
complexes were resistant to treatment with reducing agents in vivo,
indicating that the presence and stability of the disulfide interaction
between gp70 and Pr15E are not dependent on the cellular redox state. However, disulfide-bonded Env complexes were disrupted in lysates of
nonalkylated cells in a time-, temperature-, and pH-dependent fashion.
Disruption seemed not to be caused by a cellular factor but is probably
due to a thiol-disulfide exchange reaction occurring within the Env
complex after solubilization. The possibility that alkylating agents
induce the formation of the intersubunit disulfide linkage was excluded
by showing that disulfide-linked gp70-Pr15E complexes exist in freshly
made lysates of nonalkylated cells and that disruption of the complexes
can be prevented by lowering the pH. Together, these data establish
that gp70 and Pr15E form a stable disulfide-linked complex in vivo.
*
Corresponding author. Mailing address: Department for
Biosciences at Novum, Karolinska Institute, S-141 57 Huddinge, Sweden. Phone: 46-8-608 91 21. Fax: 46-8-774 55 38. E-mail:
Dirk-Jan.Opstelten{at}cbt.ki.se.
J Virol, August 1998, p. 6537-6545, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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