1stb: Difference between revisions

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ACCOMMODATION OF INSERTION MUTATIONS ON THE SURFACE AND IN THE INTERIOR OF STAPHYLOCOCCAL NUCLEASE

OverviewOverview

Alignment of homologous amino acid sequences reveals that insertion mutations are fairly common in evolution. Hitherto, the structural consequences of insertion mutations on the surface and in the interior of proteins of known structures have received little attention. We report here the high-resolution X-ray crystal structures of 2 site-directed insertion mutants of staphylococcal nuclease. The structure of the first insertion mutant, in which 2 glycine residues were inserted on the protein surface in the amino-terminal beta-strand, has been solved to 1.70 A resolution and refined to a crystallographic R value of 0.182. The inserted residues are accommodated in a special 3-residue beta-bulge. A bridging water molecule in the newly created cavity satisfies the hydrogen bonding requirements of the beta-sheet by forming a bifurcated hydrogen bond to 1 beta-strand, and a single hydrogen bond to the other beta-strand. The second insertion mutant contains a single leucine residue inserted at the end of the third beta-strand. The structure was solved to 2.0 A resolution and refined to a final R value of 0.196. The insertion is accommodated in a register shift that changes the conformation of the flexible loop portion of the molecule, relaxing and widening the omega turn. This structural alteration results in changes in position and coordination of a bound calcium ion important for catalysis. These structures illustrate important differences in how amino acid insertions are accommodated: as localized bulges, and as extensive register shifts.

About this StructureAbout this Structure

1STB is a Single protein structure of sequence from Staphylococcus aureus with and as ligands. Active as Micrococcal nuclease, with EC number 3.1.31.1 Full crystallographic information is available from OCA.

ReferenceReference

Accommodation of insertion mutations on the surface and in the interior of staphylococcal nuclease., Keefe LJ, Quirk S, Gittis A, Sondek J, Lattman EE, Protein Sci. 1994 Mar;3(3):391-401. PMID:8019410

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-[[Image:1stb.jpg|left|200px]]<br /><applet load="1stb" size="450" color="white" frame="true" align="right" spinBox="true"
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 '''ACCOMMODATION OF INSERTION MUTATIONS ON THE SURFACE AND IN THE INTERIOR OF STAPHYLOCOCCAL NUCLEASE'''<br />
 ==Overview==
-Alignment of homologous amino acid sequences reveals that insertion, mutations are fairly common in evolution. Hitherto, the structural, consequences of insertion mutations on the surface and in the interior of, proteins of known structures have received little attention. We report, here the high-resolution X-ray crystal structures of 2 site-directed, insertion mutants of staphylococcal nuclease. The structure of the first, insertion mutant, in which 2 glycine residues were inserted on the protein, surface in the amino-terminal beta-strand, has been solved to 1.70 A, resolution and refined to a crystallographic R value of 0.182. The, inserted residues are accommodated in a special 3-residue beta-bulge. A, bridging water molecule in the newly created cavity satisfies the hydrogen, bonding requirements of the beta-sheet by forming a bifurcated hydrogen, bond to 1 beta-strand, and a single hydrogen bond to the other, beta-strand. The second insertion mutant contains a single leucine residue, inserted at the end of the third beta-strand. The structure was solved to, 2.0 A resolution and refined to a final R value of 0.196. The insertion is, accommodated in a register shift that changes the conformation of the, flexible loop portion of the molecule, relaxing and widening the omega, turn. This structural alteration results in changes in position and, coordination of a bound calcium ion important for catalysis. These, structures illustrate important differences in how amino acid insertions, are accommodated: as localized bulges, and as extensive register shifts.
+Alignment of homologous amino acid sequences reveals that insertion mutations are fairly common in evolution. Hitherto, the structural consequences of insertion mutations on the surface and in the interior of proteins of known structures have received little attention. We report here the high-resolution X-ray crystal structures of 2 site-directed insertion mutants of staphylococcal nuclease. The structure of the first insertion mutant, in which 2 glycine residues were inserted on the protein surface in the amino-terminal beta-strand, has been solved to 1.70 A resolution and refined to a crystallographic R value of 0.182. The inserted residues are accommodated in a special 3-residue beta-bulge. A bridging water molecule in the newly created cavity satisfies the hydrogen bonding requirements of the beta-sheet by forming a bifurcated hydrogen bond to 1 beta-strand, and a single hydrogen bond to the other beta-strand. The second insertion mutant contains a single leucine residue inserted at the end of the third beta-strand. The structure was solved to 2.0 A resolution and refined to a final R value of 0.196. The insertion is accommodated in a register shift that changes the conformation of the flexible loop portion of the molecule, relaxing and widening the omega turn. This structural alteration results in changes in position and coordination of a bound calcium ion important for catalysis. These structures illustrate important differences in how amino acid insertions are accommodated: as localized bulges, and as extensive register shifts.
 ==About this Structure==
-STB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Staphylococcus_aureus Staphylococcus aureus] with CA and THP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Micrococcal_nuclease Micrococcal nuclease], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.31.1 3.1.31.1] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1STB OCA].
+STB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Staphylococcus_aureus Staphylococcus aureus] with <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=THP:'>THP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Micrococcal_nuclease Micrococcal nuclease], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.31.1 3.1.31.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1STB OCA].
 ==Reference==
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 [[Category: Staphylococcus aureus]]
 [[Category: Gittis, A.]]
-[[Category: Keefe, L.J.]]
+[[Category: Keefe, L J.]]
-[[Category: Lattman, E.E.]]
+[[Category: Lattman, E E.]]
 [[Category: Quirk, S.]]
 [[Category: CA]]
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 [[Category: hydrolase(phosphoric diester)]]
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