Linked Life Data

8702542

Source:http://linkedlifedata.com/resource/pubmed/id/8702542

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
31
pubmed:dateCreated
1996-9-24
pubmed:abstractText
Prenyltransferases catalyze the consecutive condensation of isopentenyl diphosphate with allylic diphosphates to produce prenyl diphosphates whose chain lengths are absolutely determined by each enzyme. To investigate the mechanism of the consecutive reaction and the determination of the ultimate chain length, a random mutational approach was planned. A geranylgeranyl-diphosphate synthase gene from Sulfolobus acidocaldarius was randomly mutagenized by NaNO2 treatment to construct a library of mutated geranylgeranyl-diphosphate synthase genes on a yeast expression vector. The library was screened for suppression of a pet phenotype of yeast C296-LH3, which is deficient in hexaprenyl-diphosphate synthase. Five mutants that could grow on a YEPG plate, which contained only glycerol as an energy source instead of glucose, were selected from approximately 1,400 mutants. All selected mutated enzymes catalyzed the formation of polyprenyl diphosphates with prenyl chains longer than geranylgeranyl diphosphate. Especially mutants 1, 3, and 5 showed the strongest elongation activity to produce large amounts of geranylfarnesyl diphosphate with a concomitant amount of hexaprenyl diphosphate. Sequence analysis revealed that each mutant contained a few amino acid substitutions and that the mutation of Phe-77, which is located on the fifth amino acid upstream from the first aspartate-rich consensus motif, is the most effective for elongating the ultimate product. Amino acid alignment of known prenyltransferases around this position and our previous observations on farnesyl-diphosphate synthase (Ohnuma, S.-i., Nakazawa, T., Hemmi, H., Hallberg, A.-M., Koyama, T., Ogura, K., and Nishino, T.(1996) J. Biol. Chem. 271, 10087-10095) clearly indicate that the amino acid at the position of all prenyltransferases must regulate the chain elongation.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
2
pubmed:volume
271
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
18831-7
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8702542-Alcohols, pubmed-meshheading:8702542-Alkyl and Aryl Transferases, pubmed-meshheading:8702542-Amino Acid Sequence, pubmed-meshheading:8702542-Base Sequence, pubmed-meshheading:8702542-DNA, Bacterial, pubmed-meshheading:8702542-DNA Primers, pubmed-meshheading:8702542-Dimethylallyltranstransferase, pubmed-meshheading:8702542-Farnesyltranstransferase, pubmed-meshheading:8702542-Genes, Bacterial, pubmed-meshheading:8702542-Molecular Sequence Data, pubmed-meshheading:8702542-Mutation, pubmed-meshheading:8702542-Phenotype, pubmed-meshheading:8702542-Saccharomyces cerevisiae, pubmed-meshheading:8702542-Sequence Homology, Amino Acid, pubmed-meshheading:8702542-Substrate Specificity, pubmed-meshheading:8702542-Sulfolobus acidocaldarius, pubmed-meshheading:8702542-Transferases
pubmed:year
1996
pubmed:articleTitle
Conversion of product specificity of archaebacterial geranylgeranyl-diphosphate synthase. Identification of essential amino acid residues for chain length determination of prenyltransferase reaction.
pubmed:affiliation
Department of Biochemistry and Engineering, Tohoku University, Aoba Aramaki, Aoba-ku, Sendai 980-77, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't