Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
8
|
| pubmed:dateCreated |
1995-4-4
|
| pubmed:abstractText |
Bovine heart fructose 6-P,2-kinase:fructose 2,6-bisphosphatase was expressed in Escherichia coli. In order to determine the role of the carboxyl-terminal peptide, 49 and 78 amino acids from the C-terminus were deleted using oligonucleotide-directed mutagenesis. The expressed wild-type and mutant enzymes were purified to homogeneity, and the steady-state kinetics of the mutant enzymes were compared to those of the wild-type enzyme. Deletion of 49 residues (Del 49) resulted in a 35% decrease in KmFru6P, a 36% increase in Vmax, and a 2-fold increase in Kcat/Km of the kinase. There was no change in the kinetic properties of the phosphatase activity. Deletion of 78 residues (Del 78) resulted in a 4.5-fold decrease in KmFru6P, a 2.5-fold increase in Vmax, a 12-fold increase in kcat/Km of the kinase, and a 3-fold increase in kcat/Km of the phosphatase. Phosphorylation of the wild-type and Del 49 enzymes resulted in decreased KmFru6P and activation of the kinase without affecting the phosphatase activity. Thermal inactivation rates of the wild-type and Del 49 enzymes were similar, but the rate of Del 78 was more rapid. The phosphorylated wild-type and Del 49 enzymes were more sensitive to thermal inactivation than the dephospho forms. Urea inactivation of the kinase and phosphatase of wild-type and Del 49 were similar, but Del 78 was more sensitive to urea. All phosphorylated enzymes were more susceptible to urea inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Multienzyme Complexes,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphofructokinase-2,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoric Monoester Hydrolases,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphotransferases,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Urea
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Feb
|
| pubmed:issn |
0006-2960
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
28
|
| pubmed:volume |
34
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
2553-9
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:7873535-Amino Acid Sequence,
pubmed-meshheading:7873535-Animals,
pubmed-meshheading:7873535-Base Sequence,
pubmed-meshheading:7873535-Binding Sites,
pubmed-meshheading:7873535-Cattle,
pubmed-meshheading:7873535-Cloning, Molecular,
pubmed-meshheading:7873535-DNA, Complementary,
pubmed-meshheading:7873535-DNA Primers,
pubmed-meshheading:7873535-Escherichia coli,
pubmed-meshheading:7873535-Gene Expression,
pubmed-meshheading:7873535-Kinetics,
pubmed-meshheading:7873535-Molecular Sequence Data,
pubmed-meshheading:7873535-Multienzyme Complexes,
pubmed-meshheading:7873535-Mutagenesis, Site-Directed,
pubmed-meshheading:7873535-Myocardium,
pubmed-meshheading:7873535-Phosphofructokinase-2,
pubmed-meshheading:7873535-Phosphoric Monoester Hydrolases,
pubmed-meshheading:7873535-Phosphorylation,
pubmed-meshheading:7873535-Phosphotransferases,
pubmed-meshheading:7873535-Recombinant Proteins,
pubmed-meshheading:7873535-Sequence Deletion,
pubmed-meshheading:7873535-Urea
|
| pubmed:year |
1995
|
| pubmed:articleTitle |
Expression of bovine heart fructose 6-phosphate,2-kinase:fructose 2,6-bisphosphatase and determination of the role of the carboxyl terminus by mutagenesis.
|
| pubmed:affiliation |
Department of Veterans Affairs Medical Center, Dallas, TX 75216.
|
| pubmed:publicationType |
Journal Article,
In Vitro,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.
|