Expression and purification of functional human glycogen synthase-1 (hGYS1) in insect cells

dc.contributor.authorKhanna, May
dc.contributor.authorImasaki, Tsuyoshi
dc.contributor.authorChikwana, Vimbai M.
dc.contributor.authorPerez-Miller, Samantha
dc.contributor.authorHunter, Gerald O.
dc.contributor.authorMosley, Amber
dc.contributor.authorTakagi, Yuichiro
dc.contributor.authorHurley, Thomas D.
dc.contributor.departmentBiochemistry and Molecular Biology, School of Medicine
dc.date.accessioned2025-05-08T11:30:19Z
dc.date.available2025-05-08T11:30:19Z
dc.date.issued2013
dc.description.abstractWe have successfully expressed and purified active human glycogen synthase-1 (hGYS1). Successful production of the recombinant hGYS1 protein was achieved by co-expression of hGYS1 and rabbit glycogenin (rGYG1) using the MultiBac baculovirus expression system (BEVS). Functional measurements of activity ratios of hGYS1 in the absence and presence of glucose-6-phosphate and treatment with phosphatase indicate that the expressed protein is heavily phosphorylated. We used mass spectrometry to further characterize the sites of phosphorylation, which include most of the known regulatory phosphorylation sites, as well as several sites unique to the insect cell over-expression. Obtaining large quantities of functional hGYS1 will be invaluable for future structural studies as well as detailed studies on the effects on specific sites of phosphorylation.
dc.eprint.versionAuthor's manuscript
dc.identifier.citationKhanna M, Imasaki T, Chikwana VM, et al. Expression and purification of functional human glycogen synthase-1 (hGYS1) in insect cells. Protein Expr Purif. 2013;90(2):78-83. doi:10.1016/j.pep.2013.05.007
dc.identifier.urihttps://hdl.handle.net/1805/47881
dc.language.isoen_US
dc.publisherElsevier
dc.relation.isversionof10.1016/j.pep.2013.05.007
dc.relation.journalProtein Expression and Purification
dc.rightsPublisher Policy
dc.sourcePMC
dc.subjectHuman glycogen synthase-1
dc.subjectMass spectrometry
dc.subjectMultiBac baculovirus expression system
dc.subjectPhosphorylation
dc.titleExpression and purification of functional human glycogen synthase-1 (hGYS1) in insect cells
dc.typeArticle
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