2i6r: Difference between revisions

Revision as of 09:17, 13 February 2008

Crystal structure of E. coli HypE, a hydrogenase maturation protein

OverviewOverview

Hydrogenases are enzymes involved in hydrogen metabolism, utilizing H2 as, an electron source. [NiFe] hydrogenases are heterodimeric Fe-S proteins, with a large subunit containing the reaction centre involving Fe and Ni, metal ions and a small subunit containing one or more Fe-S clusters., Maturation of the [NiFe]-hydrogenase involves assembly of, non-proteinaceous ligands on the large subunit by accessory proteins, encoded by the hyp operon. HypE is an essential accessory protein and, participates in the synthesis of two cyano groups found in the large, subunit. We report the crystal structure of Escherichia coli HypE at 2.0 A, resolution. HypE exhibits fold similar to PurM and ThiL and forms dimers., The C-terminal catalytically-essential Cys336 is internalized at the dimer, interface between the N- and C-terminal domains. A mechanism for, dehydration of the thiocarbamate to the thiocyanate is proposed, involving, Asp83 and Glu272. The interactions of HypE and HypF were characterized in, detail by surface plasmon resonance and isothermal titration calorimetry, revealing a Kd for their association of approximately 400 nM. The, stoichiometry and molecular weights of the complex were verified by size, exclusion chromatography and gel scanning densitometry. These experiments, reveal that HypE and HypF associate to form a stoichiometric, hetero-oligomeric complex predominantly consisting of a [EF]2, heterotetramer which exists in a dynamic equilibrium with the EF, heterodimer. The surface plasmon resonance results indicate that a, conformational change occurs upon heterodimerization which facilitates, formation of a productive complex as part of the carbamate transfer, reaction.

About this StructureAbout this Structure

2I6R is a Single protein structure of sequence from Escherichia coli o157:h7. Full crystallographic information is available from OCA.

ReferenceReference

Structure of [NiFe]-Hydrogenase Maturation Protein HypE from Escherichia coli and its Interaction with HypF., Rangarajan ES, Asinas A, Proteau A, Munger C, Baardsnes J, Iannuzzi P, Matte A, Cygler M, J Bacteriol. 2007 Dec 7;. PMID:18065529

Page seeded by OCA on Wed Feb 13 08:17:35 2008

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

OCA

@@ Line 2: / Line 2: @@
 caption="2i6r, resolution 2.51&Aring;" />
 '''Crystal structure of E. coli HypE, a hydrogenase maturation protein'''<br />
+==Overview==
+Hydrogenases are enzymes involved in hydrogen metabolism, utilizing H2 as, an electron source. [NiFe] hydrogenases are heterodimeric Fe-S proteins, with a large subunit containing the reaction centre involving Fe and Ni, metal ions and a small subunit containing one or more Fe-S clusters., Maturation of the [NiFe]-hydrogenase involves assembly of, non-proteinaceous ligands on the large subunit by accessory proteins, encoded by the hyp operon. HypE is an essential accessory protein and, participates in the synthesis of two cyano groups found in the large, subunit. We report the crystal structure of Escherichia coli HypE at 2.0 A, resolution. HypE exhibits fold similar to PurM and ThiL and forms dimers., The C-terminal catalytically-essential Cys336 is internalized at the dimer, interface between the N- and C-terminal domains. A mechanism for, dehydration of the thiocarbamate to the thiocyanate is proposed, involving, Asp83 and Glu272. The interactions of HypE and HypF were characterized in, detail by surface plasmon resonance and isothermal titration calorimetry, revealing a Kd for their association of approximately 400 nM. The, stoichiometry and molecular weights of the complex were verified by size, exclusion chromatography and gel scanning densitometry. These experiments, reveal that HypE and HypF associate to form a stoichiometric, hetero-oligomeric complex predominantly consisting of a [EF]2, heterotetramer which exists in a dynamic equilibrium with the EF, heterodimer. The surface plasmon resonance results indicate that a, conformational change occurs upon heterodimerization which facilitates, formation of a productive complex as part of the carbamate transfer, reaction.
 ==About this Structure==
 I6R is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_o157:h7 Escherichia coli o157:h7]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2I6R OCA].
+==Reference==
+Structure of [NiFe]-Hydrogenase Maturation Protein HypE from Escherichia coli and its Interaction with HypF., Rangarajan ES, Asinas A, Proteau A, Munger C, Baardsnes J, Iannuzzi P, Matte A, Cygler M, J Bacteriol. 2007 Dec 7;. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18065529 18065529]
 [[Category: Escherichia coli o157:h7]]
 [[Category: Single protein]]
@@ Line 20: / Line 26: @@
 [[Category: unknown function]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 12:41:59 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Feb 13 08:17:35 2008''