TY - CHAP
T1 - Structural and biochemical analysis of a bacterial glycosyltransferase
AU - Zhu, Fan
AU - Wu, Ren
AU - Zhang, Hua
AU - Wu, Hui
PY - 2013
Y1 - 2013
N2 - Glycosyltransferases (GTs) are a large family of enzymes that specifically transfer sugar moieties to a diverse range of substrates. The process of bacterial glycosylation (such as biosynthesis of glycolipids, glycoproteins, and polysaccharides) has been studied extensively, yet the majority of GTs involved remains poorly characterized. Besides predicting enzymatic parameters of GTs, the resolution of three-dimensional structures of GTs can help to determine activity, donor sugar binding, and acceptor substrate binding sites. It also facilitates amino acid sequence-based structural modeling and biochemical characterization of their homologues. Here we describe a general procedure to accomplish expression and purification of soluble and active recombinant GTs. Enzymatic characterization, and crystallization of GTs, and data refinement for structural analysis are also covered in this protocol.
AB - Glycosyltransferases (GTs) are a large family of enzymes that specifically transfer sugar moieties to a diverse range of substrates. The process of bacterial glycosylation (such as biosynthesis of glycolipids, glycoproteins, and polysaccharides) has been studied extensively, yet the majority of GTs involved remains poorly characterized. Besides predicting enzymatic parameters of GTs, the resolution of three-dimensional structures of GTs can help to determine activity, donor sugar binding, and acceptor substrate binding sites. It also facilitates amino acid sequence-based structural modeling and biochemical characterization of their homologues. Here we describe a general procedure to accomplish expression and purification of soluble and active recombinant GTs. Enzymatic characterization, and crystallization of GTs, and data refinement for structural analysis are also covered in this protocol.
KW - Crystallization and data refinement
KW - Glycosyltransferase assays
KW - Glycosyltransferases
KW - Protein purification
UR - http://www.scopus.com/inward/record.url?scp=84880467265&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84880467265&partnerID=8YFLogxK
U2 - 10.1007/978-1-62703-465-4_3
DO - 10.1007/978-1-62703-465-4_3
M3 - Chapter
C2 - 23765651
AN - SCOPUS:84880467265
SN - 9781627034647
T3 - Methods in Molecular Biology
SP - 29
EP - 39
BT - Glycosyltransferases
PB - Humana Press Inc.
ER -