TY - CHAP
T1 - Structural Determination of Glucosyltransferase C by Cryo-Electron Microscopy
AU - Xie, Qing
AU - Cao, Jianhao
AU - Zhang, Hua
AU - Wu, Hui
N1 - Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. 2024.
PY - 2024
Y1 - 2024
N2 - Biofilm formation is a critical factor in the development of cariogenic virulence of Streptococcus mutans. S. mutans has evolved a concerted mechanism to synthesize a biofilm matrix from dietary sugars by a family of glucosyltransferases (Gtfs). Three Gtfs, GtfB, C, and D utilize sucrose to form a sticky polymer consisting of insoluble and soluble glucans, a biofilm foundation. Each Gtf possesses two distinct domains, an N-terminal enzyme catalytic domain and a C-terminal glucan binding domain. X-ray crystallographic studies have determined a three-dimensional structure of the catalytic domain of GtfC, however, the structure of the C-terminal domain and the overall structure of Gtfs are unknown. Here, we provided a protocol that will guide us to solve Gtf structures using cryo-electron microscopy (cryo-EM). Cryo-EM and X-ray crystallography are two widely used techniques for determining protein structures, and both methods have their own advantages and limitations. Additionally, we also predicted the full-length GtfC structural using AlphaFold2. Overall, the combination of AlphaFold 2 with experimental approaches offers a powerful and synergistic strategy for protein structure determination, accelerating the pace of scientific discovery and enabling new insights into the molecular basis that govern the biosynthetic activities of Gtfs, which may inform the design of more effective treatments for cariogenic biofilms and associated diseases.
AB - Biofilm formation is a critical factor in the development of cariogenic virulence of Streptococcus mutans. S. mutans has evolved a concerted mechanism to synthesize a biofilm matrix from dietary sugars by a family of glucosyltransferases (Gtfs). Three Gtfs, GtfB, C, and D utilize sucrose to form a sticky polymer consisting of insoluble and soluble glucans, a biofilm foundation. Each Gtf possesses two distinct domains, an N-terminal enzyme catalytic domain and a C-terminal glucan binding domain. X-ray crystallographic studies have determined a three-dimensional structure of the catalytic domain of GtfC, however, the structure of the C-terminal domain and the overall structure of Gtfs are unknown. Here, we provided a protocol that will guide us to solve Gtf structures using cryo-electron microscopy (cryo-EM). Cryo-EM and X-ray crystallography are two widely used techniques for determining protein structures, and both methods have their own advantages and limitations. Additionally, we also predicted the full-length GtfC structural using AlphaFold2. Overall, the combination of AlphaFold 2 with experimental approaches offers a powerful and synergistic strategy for protein structure determination, accelerating the pace of scientific discovery and enabling new insights into the molecular basis that govern the biosynthetic activities of Gtfs, which may inform the design of more effective treatments for cariogenic biofilms and associated diseases.
KW - Alpha fold
KW - Cryo-electron microscopy
KW - Glucosyltransferase
KW - Streptococcus mutans
KW - Structural determination
UR - http://www.scopus.com/inward/record.url?scp=85174899690&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85174899690&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-3491-2_18
DO - 10.1007/978-1-0716-3491-2_18
M3 - Chapter
C2 - 37815721
AN - SCOPUS:85174899690
T3 - Methods in Molecular Biology
SP - 227
EP - 237
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -