Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
|
pubmed:dateCreated |
1996-7-12
|
pubmed:abstractText |
Selected physical and thermodynamic parameters for Escherichia coli alanyl-tRNA synthetase (AlaRS) have been determined primarily to assess the quaternary structure of this enzyme. The extinction coefficient (epsilon) at 280 nm was determined experimentally to be 0.71 ml mg-1 cm-1, and the partial specific volume (nu) was calculated from the amino acid composition to be 0.73 ml g-1. From viscosity experiments the intrinsic viscosity (eta) of AlaRS was extrapolated to be 3.4 ml g-1 and the degree of hydration (delta 1) estimated to be 0.67 gH2O g(-1)(AlaRS). Laser light-scattering studies indicated some heterogeneity; a radius of 6.3 nm was calculated for the major fraction with a diffusion coefficient (D20,W) of 3.89 x 10(-7) cm2 s-1. In 50 mM Hepes, pH 7.5, 20 mM KCl, 2 mM 2-mercaptoethanol and at a protein concentration of 4.2 mg ml-1 the sedimentation coefficient (S20,W) was 6.36 S; this value increased slightly when the protein concentration was decreased. The combination of S20,W and D20,W under these conditions yielded a molecular weight of approximately 186,000 Da, corresponding to a dimer. The S20,W was virtually independent of temperature in the range of 10-37 degrees C, while an Arrhenius plot of aminoacylation activity was biphasic. The isoelectric point was determined experimentally to be 4.9. Sedimentation equilibrium data were best fit to a decamer association complex in which dimeric AlaRS is the predominant species at 25 degrees C.
|
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Apr
|
pubmed:issn |
0003-9861
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
15
|
pubmed:volume |
328
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
295-301
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:8645007-Alanine-tRNA Ligase,
pubmed-meshheading:8645007-Escherichia coli,
pubmed-meshheading:8645007-Hydrogen-Ion Concentration,
pubmed-meshheading:8645007-Isoelectric Focusing,
pubmed-meshheading:8645007-Kinetics,
pubmed-meshheading:8645007-Lasers,
pubmed-meshheading:8645007-Light,
pubmed-meshheading:8645007-Macromolecular Substances,
pubmed-meshheading:8645007-Molecular Weight,
pubmed-meshheading:8645007-Recombinant Proteins,
pubmed-meshheading:8645007-Scattering, Radiation,
pubmed-meshheading:8645007-Thermodynamics,
pubmed-meshheading:8645007-Viscosity
|
pubmed:year |
1996
|
pubmed:articleTitle |
Further characterization of Escherichia coli alanyl-tRNA synthetase.
|
pubmed:affiliation |
Department of Biochemistry, Loma Linda University School of Medicine, California 92350-0001, USA.
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.
|