1E2B

NMR STRUCTURE OF THE C10S MUTANT OF ENZYME IIB CELLOBIOSE OF THE PHOSPHOENOL-PYRUVATE DEPENDENT PHOSPHOTRANSFERASE SYSTEM OF ESCHERICHIA COLI, 17 STRUCTURES

1E2B の概要

• エントリーDOI: 10.2210/pdb1e2b/pdb
• 分子名称: ENZYME IIB-CELLOBIOSE (1 entity in total)
• 機能のキーワード: enzyme iib-cellobiose, phosphotransferase system, transferase, sugar transport, phosphorylation
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cytoplasm: P69795
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 11423.45

構造登録者

Ab, E.,Schuurman-Wolters, G.,Reizer, J.,Saier, M.H.,Dijkstra, K.,Scheek, R.M.,Robillard, G.T. (登録日: 1996-11-15, 公開日: 1997-07-23, 最終更新日: 2024-05-22)

主引用文献

Ab, E.,Schuurman-Wolters, G.,Reizer, J.,Saier, M.H.,Dijkstra, K.,Scheek, R.M.,Robillard, G.T.
The NMR side-chain assignments and solution structure of enzyme IIBcellobiose of the phosphoenolpyruvate-dependent phosphotransferase system of Escherichia coli.
Protein Sci., 6:304-314, 1997

PubMed Abstract: The assignment of the side-chain NMR resonances and the determination of the three-dimensional solution structure of the C10S mutant of enzyme IIBcellobiose (IIBcel) of the phosphoenolpyruvate-dependent phosphotransferase system of Escherichia coli are presented. The side-chain resonances were assigned nearly completely using a variety of mostly heteronuclear NMR experiments, including HCCH-TOCSY, HCCH-COSY, and COCCH-TOCSY experiments as well as CBCACOHA, CBCA(CO)NH, and HBHA(CBCA)(CO)NH experiments. In order to obtain the three-dimensional structure, NOE data were collected from 15N-NOESY-HSQC, 13C-HSQC-NOESY, and 2D NOE experiments. The distance restraints derived from these NOE data were used in distance geometry calculations followed by molecular dynamics and simulated annealing protocols. In an iterative procedure, additional NOE assignments were derived from the calculated structures and new structures were calculated. The final set of structures, calculated with approximately 2000 unambiguous and ambiguous distance restraints, has an rms deviation of 1.1 A on C alpha atoms. IIBcel consists of a four stranded parallel beta-sheet, in the order 2134. The sheet is flanked with two and three alpha-helices on either side. Residue 10, a cysteine in the wild-type enzyme, which is phosphorylated during the catalytic cycle, is located at the end of the first beta-strand. A loop that is proposed to be involved in the binding of the phosphoryl-group follows the cysteine. The loop appears to be disordered in the unphosphorylated state.

PubMed: 9041631

実験手法

SOLUTION NMR