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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
22
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pubmed:dateCreated |
1986-12-17
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pubmed:abstractText |
Gene 32 protein (g32P) isolated from bacteriophage T4-infected Escherichia coli and from an overproduction vector derived from the plasmid pKC30 contains 1 mol of tightly incorporated Zn(II) per mol of protein. A linear incorporation of three molar equivalents of p-hydroxymercuriphenylsulfonate (PMPS) results in a linear release of 1.1 mol of Zn(II) from the protein. Reversal of formation of the g32P-PMPS complex with thiol in the presence of EDTA results in a zinc-free apo-g32P. Cd(II) and Co(II) can be exchanged with the intrinsic Zn(II) ion. The Cd(II) protein shows a charge-transfer band at approximately 250 nm. The Co(II) protein shows a set of absorption bands typical of a tetrahedral Co(II) complex (epsilon max = 660 M-1 X cm-1 at 645 nm), and two intense charge-transfer bands are present at 355 nm (epsilon = 2,250 M-1 X cm-1) and 320 nm (epsilon = 3,175 M-1 X cm-1). These observations are consistent with three cysteines as ligands to the Zn(II) ion in g32P. Zn(II) g32P undergoes precise limited proteolysis by trypsin to produce the small fragments A and B and the core, g32P-(A + B). Under identical conditions, apo-g32P is hydrolyzed rapidly beyond the g32P-(A + B) stage to produce many proteolyzed fragments. Fluorescence quenching experiments show that at low protein concentration apo-g32P has markedly altered binding affinity for poly(dT) relative to native g32P. Three of the four cysteines of g32P are found in a tyrosine-rich sequence corresponding to residues 72-116 and implicated in DNA binding by 1H NMR investigations. Zn(II) appears to provide a conformational element contributing to DNA binding by coordinating the cysteine and possibly histidine side chains in the sequence -Cys-X3-His-X5-Cys-X2-Cys-, residues 77-90, located in the DNA binding domain of g32P.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-100782,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-1090613,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-1125177,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-212428,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-226522,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-2421409,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3013293,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3082355,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3094579,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3486003,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3514922,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-3515197,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-353052,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-353053,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-4040853,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-4115107,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-4608147,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-488110,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-5455134,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6196359,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6243083,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6257686,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6303852,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-632279,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6367821,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6371808,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6389552,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-6974567,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3490667-791945
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
83
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
8452-6
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:3490667-Amino Acid Sequence,
pubmed-meshheading:3490667-Cadmium,
pubmed-meshheading:3490667-Cobalt,
pubmed-meshheading:3490667-DNA, Single-Stranded,
pubmed-meshheading:3490667-DNA-Binding Proteins,
pubmed-meshheading:3490667-Metalloproteins,
pubmed-meshheading:3490667-Poly T,
pubmed-meshheading:3490667-T-Phages,
pubmed-meshheading:3490667-Trypsin,
pubmed-meshheading:3490667-Viral Proteins,
pubmed-meshheading:3490667-Zinc
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pubmed:year |
1986
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pubmed:articleTitle |
Gene 32 protein, the single-stranded DNA binding protein from bacteriophage T4, is a zinc metalloprotein.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.
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