HIV TAT variants differentially influence the production of glucocerebrosidase in Sf9 cells.
Gaucher disease, the most common lysosomal storage disorder, is currently treated with enzyme replacement therapy. This approach, however, is ineffective in altering the progression of neurodegeneration in type 2 and type 3 patients due to the difficulty of transferring the recombinant enzyme across the blood-brain barrier. Human immunodeficiency virus type 1 trans-activating transcriptional activator protein (HIV TAT) contains a protein transduction domain that can be added to a fusion protein partner to allow for transport of the partner across membranes. Consequently, we examined the creation, production, and secretion of fusion constructs containing glucocerebrosidase and either wild-type TAT or modified TAT in Sf9 cells. All three constructs exhibited successful expression, with wild-type TAT chimeras showing lower levels of expression than modified TAT chimeras.
Vaags
Andrea K
AK
Department of Biology, University of Victoria, Victoria, BC, Canada.
Campbell
Tessa N
TN
Choy
Francis Y M
FY
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005776:Gaucher Disease / Q000378:metabolism / Q000628:therapy; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism*; D005962:Glucosylceramidase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D021381:Protein Transport / Q000235:genetics; D014158:Transcription, Genetic; D014161:Transduction, Genetic
D015696:Gene Products, tat; D005962:Glucosylceramidase