1e2b: Difference between revisions

Revision as of 13:23, 21 February 2008

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

OverviewOverview

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.

About this StructureAbout this Structure

1E2B is a Single protein structure of sequence from Escherichia coli. Active as Protein-N(pi)-phosphohistidine--sugar phosphotransferase, with EC number 2.7.1.69 Full crystallographic information is available from OCA.

ReferenceReference

The NMR side-chain assignments and solution structure of enzyme IIBcellobiose of the phosphoenolpyruvate-dependent phosphotransferase system of Escherichia coli., Ab E, Schuurman-Wolters G, Reizer J, Saier MH, Dijkstra K, Scheek RM, Robillard GT, Protein Sci. 1997 Feb;6(2):304-14. PMID:9041631

Page seeded by OCA on Thu Feb 21 12:23:01 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 1: / Line 1: @@
-[[Image:1e2b.gif|left|200px]]<br /><applet load="1e2b" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1e2b.gif|left|200px]]<br /><applet load="1e2b" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1e2b" />
 '''NMR STRUCTURE OF THE C10S MUTANT OF ENZYME IIB CELLOBIOSE OF THE PHOSPHOENOL-PYRUVATE DEPENDENT PHOSPHOTRANSFERASE SYSTEM OF ESCHERICHIA COLI, 17 STRUCTURES'''<br />
 ==Overview==
-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.
+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.
 ==About this Structure==
-E2B is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Active as [http://en.wikipedia.org/wiki/Protein-N(pi)-phosphohistidine--sugar_phosphotransferase Protein-N(pi)-phosphohistidine--sugar phosphotransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.69 2.7.1.69] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1E2B OCA].
+E2B is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Active as [http://en.wikipedia.org/wiki/Protein-N(pi)-phosphohistidine--sugar_phosphotransferase Protein-N(pi)-phosphohistidine--sugar phosphotransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.69 2.7.1.69] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1E2B OCA].
 ==Reference==
@@ Line 17: / Line 17: @@
 [[Category: Dijkstra, K.]]
 [[Category: Reizer, J.]]
-[[Category: Robillard, G.T.]]
+[[Category: Robillard, G T.]]
-[[Category: Saier, M.H.]]
+[[Category: Saier, M H.]]
-[[Category: Scheek, R.M.]]
+[[Category: Scheek, R M.]]
 [[Category: Schuurman-Wolters, G.]]
 [[Category: enzyme iib-cellobiose]]
@@ Line 27: / Line 27: @@
 [[Category: transferase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 13:44:36 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:23:01 2008''