Linked Life Data

11669634

Source:http://linkedlifedata.com/resource/pubmed/id/11669634

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
43
pubmed:dateCreated
2001-10-23
pubmed:abstractText
Human polynucleotide kinase (hPNK) is a putative DNA repair enzyme in the base excision repair pathway required for processing and rejoining strand-break termini. This study represents the first systematic examination of the physical properties of this enzyme. The protein was produced in Escherichia coli as a His-tagged protein, and the purified recombinant protein exhibited both the kinase and the phosphatase activities. The predicted relative molecular mass (M(r)) of the 521 amino acid polypeptide encoded by the sequenced cDNA for PNK and the additional 21 amino acids of the His tag is 59,538. The M(r) determined by low-speed sedimentation equilibrium under nondenaturing conditions was 59,600 +/- 1000, indicating that the protein exists as a monomer, in contrast to T4 phage PNK, which exists as a homotetramer. The size and shape of hPNK in solution were determined by analytical ultracentrifugation studies. The protein was found to have an intrinsic sedimentation coefficient, s(0)(20,w), of 3.54 S and a Stokes radius, R(s), of 37.5 A. These hydrodynamic data, together with the M(r) of 59 600, suggest that hPNK is a moderately asymmetric protein with an axial ratio of 5.51. Analysis of the secondary structure of hPNK on the basis of circular dichroism spectra, which revealed the presence of two negative dichroic bands located at 218 and 209 nm, with ellipticity values of -7200 +/- 300 and -7800 +/- 300 deg x cm(2) x d(mol(-1), respectively, indicated the presence of approximately 50% beta-structure and 25% alpha-helix. Binding of ATP to the protein induced an increase in beta-structure and perturbed tryptophan, tyrosine, and phenylalanine signals observed by aromatic CD and UV difference spectroscopy.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
30
pubmed:volume
40
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
12967-73
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11669634-Adenosine Triphosphate, pubmed-meshheading:11669634-Amino Acids, pubmed-meshheading:11669634-Animals, pubmed-meshheading:11669634-Circular Dichroism, pubmed-meshheading:11669634-Cloning, Molecular, pubmed-meshheading:11669634-DNA, Complementary, pubmed-meshheading:11669634-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:11669634-Escherichia coli, pubmed-meshheading:11669634-Humans, pubmed-meshheading:11669634-Phenylalanine, pubmed-meshheading:11669634-Polymerase Chain Reaction, pubmed-meshheading:11669634-Polynucleotide 5'-Hydroxyl-Kinase, pubmed-meshheading:11669634-Protein Binding, pubmed-meshheading:11669634-Protein Conformation, pubmed-meshheading:11669634-Protein Structure, Secondary, pubmed-meshheading:11669634-Recombinant Proteins, pubmed-meshheading:11669634-Tryptophan, pubmed-meshheading:11669634-Tyrosine, pubmed-meshheading:11669634-Ultracentrifugation, pubmed-meshheading:11669634-Ultraviolet Rays, pubmed-meshheading:11669634-Water
pubmed:year
2001
pubmed:articleTitle
Physical properties of human polynucleotide kinase: hydrodynamic and spectroscopic studies.
pubmed:affiliation
Experimental Oncology, Cross Cancer Institute, and Department of Oncology, University of Alberta, Edmonton, Alberta T6G 1Z2, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't