Membrane transplantation to correct integral membrane protein defects.
In this report we show that the tendency of certain viruses to carry host membrane proteins in their envelopes can be harnessed for transplantation of small patches of plasma membrane, including fully functional, polytopic ion channel proteins and their regulatory binding partners. As a stringent model we tested the topologically complex epithelial ion channel CFTR. Initially an attenuated vaccinia virus was found capable of transferring CFTR in a properly folded, functional and regulatable form to CFTR negative cells. Next we generated viruslike particles (VLPs) composed of retroviral structural proteins that assemble and bud at the host cell plasma membrane. These particles were also shown to mediate functional ion channel transfer. By testing the capacity of complex membrane proteins to incorporate into viral envelopes these experiments provide new insight into the permissiveness of viral envelopment, including the ability of incorporated proteins to retain function and repair defects at the cell surface, and serve as a platform for studies of ion channel and membrane protein biochemistry.
Curlee
Kimberly V
KV
Department of Human Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Hong
Jeong S
JS
Clancy
J P
JP
King
Scott A
SA
Hunter
Eric
E
Berdiev
Bakhrom
B
Benos
Dale
D
Sommerfelt
Maja A
MA
Sorscher
Eric J
EJ
Sakalian
Michael
M
D000818:Animals; D000911:Antibodies, Monoclonal; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001692:Biological Transport; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D015683:Gene Products, gag / Q000378:metabolism; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D014158:Transcription, Genetic; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics / Q000502:physiology*
D000911:Antibodies, Monoclonal; C505032:CFTR protein, human; D015683:Gene Products, gag; D008051:Lipid Bilayers; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator