6JOY

The X-ray Crystallographic Structure of Branching Enzyme from Rhodothermus obamensis STB05

Summary for 6JOY

• Entry DOI: 10.2210/pdb6joy/pdb
• Descriptor: 1,4-alpha-glucan branching enzyme GlgB (2 entities in total)
• Functional Keywords: branching enzyme, carbohydrate
• Biological source: Rhodothermus marinus (Rhodothermus obamensis)
• Total number of polymer chains: 1
• Total formula weight: 72325.02

Authors

Li, Z.F.,Ban, X.F.,Jiang, H.M.,Wang, Z.,Jin, T.C.,Li, C.M.,Gu, Z.B. (deposition date: 2019-03-25, release date: 2020-03-04, Last modification date: 2024-03-27)

Primary citation

Jiang, H.,Xie, X.,Ban, X.,Gu, Z.,Cheng, L.,Hong, Y.,Li, C.,Li, Z.
Flexible Loop in Carbohydrate-Binding Module 48 Allosterically Modulates Substrate Binding of the 1,4-alpha-Glucan Branching Enzyme.
J.Agric.Food Chem., 69:5755-5763, 2021

PubMed Abstract: The 1,4-α-glucan branching enzyme (GBE, EC 2.4.1.18) catalyzes the formation of α-1,6 branching points in starch and plays a key role in synthesis. To obtain mechanistic insights into the catalytic action of the enzyme, we first determined the crystal structure of GBE from STB05 (RoGBE) to a resolution of 2.39 Å (PDB ID: 6JOY). The structure consists of three domains: domain A, domain C, and the carbohydrate-binding module 48 (CBM48). An engineered truncated mutant lacking the CBM48 domain (ΔCBM48) showed significantly reduced ligand binding affinity and enzyme activity. Comparison of the structures of RoGBE with other GBEs showed that CBM48 of RoGBE had a longer flexible loop. Truncation of the flexible loops resulted in reduced binding affinity and activity, thereby substantiating the importance of the optimum loop structure for catalysis. In essence, our study shows that CBM48, especially the flexible loop, plays an important role in substrate binding and enzymatic activity of RoGBE. Further, based on the structural analysis, kinetics, and activity assays on wild type and mutants, as well as homology modeling, we proposed a mechanistic model (called the "lid model") to illustrate how the flexible loop triggers substrate binding, ultimately leading to catalysis.

PubMed: 33988022
DOI: 10.1021/acs.jafc.1c00293

Experimental method

X-RAY DIFFRACTION (2.392 Å)