title: Combining carbochips and mass spectrometry to study the donor specificity for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase LgtA.
abstract: A library of 11 UDP-N-acetylglucosamine analogs were rapidly screened for their activities as donors for the Neisseria meningitidis β1,3-N-acetylglucosaminyltransferase (LgtA) by direct on-chip reaction and detection with SAMDI-TOF mass spectrometry. Six of the analogs were active in this assay and were analyzed by SAMDI to characterize the kinetics toward LgtA. The analysis revealed that substitutions on C-2, C-4, and C-6 affect the activity of the donors, with bulky groups at these positions decreasing affinity of the donors for the enzyme, and also revealed that activity is strongly affected by the stereochemistry at C-3, but not C-4, of the donor. The study is also significant because it demonstrates that SAMDI can be used to both profile glycosyltransferase activities and to provide a quantitative assessment of enzyme activity.
authors.affiliation: National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, PR China.
authors.forename: Wanyi
authors.initials: W
authors.lastname: Guan
authors.forename: Lan
authors.initials: L
authors.lastname: Ban
authors.forename: Li
authors.initials: L
authors.lastname: Cai
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Wenlan
authors.initials: W
authors.lastname: Chen
authors.forename: Xianwei
authors.initials: X
authors.lastname: Liu
authors.forename: Milan
authors.initials: M
authors.lastname: Mrksich
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D008904:Miniaturization; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; C401409:LgtA protein, bacteria; D017351:N-Acetylglucosaminyltransferases