title: In vitro folding of β-1,4galactosyltransferase and polypeptide-α-N-acetylgalactosaminyltransferase from the inclusion bodies.
abstract: The aim of this article is to present a unique in vitro folding technique for glycosyltransferases to generate active proteins that can be used for X-ray crystallographic and bioconjugation protocols. Although a number of in vitro refolding methods are available, β1,4galactosyltransferases in large quantities can be made using the current protocol only. This technique is not only limited to glycosyltransferases alone but has been successfully used to refold single-chain antibodies and other molecules. Although this in vitro folding method is quite similar to other methods, it differs from them by the use of S-sulfonation of the inclusion bodies before setting up the in vitro refolding of the protein.
authors.affiliation: Structural Glycobiology Section and Basic Science Program, SAIC-Frederick, Inc., Center for Cancer Research Nanobiology Program, Center for Cancer Research, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
authors.forename: Boopathy
authors.initials: B
authors.lastname: Ramakrishnan
authors.forename: Pradman K
authors.initials: PK
authors.lastname: Qasba
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D017350:N-Acetylgalactosaminyltransferases / Q000737:chemistry*; D009237:N-Acetyllactosamine Synthase / Q000737:chemistry* / Q000302:isolation & purification; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013450:Sulfones / Q000737:chemistry; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D017350:N-Acetylgalactosaminyltransferases; D009237:N-Acetyllactosamine Synthase; D011994:Recombinant Proteins; D013450:Sulfones; D000097763:Polypeptide N-acetylgalactosaminyltransferase