Linked Life Data

8995312

Source:http://linkedlifedata.com/resource/pubmed/id/8995312

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1997-2-18
pubmed:abstractText
Protein farnesyltransferase (FTase), a heterodimer enzyme consisting of alpha and beta subunits, catalyzes the addition of farnesyl groups to the C termini of proteins such as Ras. In this paper, we report that the protein substrate specificity of yeast FTase can be switched to that of a closely related enzyme, geranylgeranyltransferase type I (GGTase I) by a single amino acid change at one of the three residues: Ser-159, Tyr-362, or Tyr-366 of its beta-subunit, Dpr1. All three Dpr1 mutants can function as either FTase or GGTase I beta subunit in vivo, although some differences in efficiency were observed. These results point to the importance of two distinct regions (one at 159 and the other at 362 and 366) of Dpr1 for the recognition of the protein substrate. Analysis of the protein, after site directed mutagenesis was used to change Ser-159 to all possible amino acids, showed that either asparagine or aspartic acid at this position allowed FTase beta to function as GGTase I beta. A similar site-directed mutagenesis study on Tyr-362 showed that leucine, methionine, or isoleucine at this position also resulted in the ability of mutant FTase beta to function as GGTase I beta. Interestingly, in both position 159 and 362 substitutions, amino acids that could change the protein substrate specificity had similar van der Waals volumes. Biochemical characterization of the S159N and Y362L mutant proteins showed that their kcat/Km values for GGTase I substrate are increased about 20-fold compared with that of the wild type protein. These results demonstrate that the conversion of the protein substrate specificity of FTase to that of GGTase I can be accomplished by introducing a distinct size amino acid at either of the two residues, 159 and 362.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
3
pubmed:volume
272
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
680-7
pubmed:dateRevised
2009-7-22
pubmed:meshHeading
pubmed-meshheading:8995312-Alkyl and Aryl Transferases, pubmed-meshheading:8995312-Amino Acid Sequence, pubmed-meshheading:8995312-Binding Sites, pubmed-meshheading:8995312-Chitin Synthase, pubmed-meshheading:8995312-Farnesyltranstransferase, pubmed-meshheading:8995312-Fungal Proteins, pubmed-meshheading:8995312-Geranyltranstransferase, pubmed-meshheading:8995312-Molecular Sequence Data, pubmed-meshheading:8995312-Mutagenesis, Site-Directed, pubmed-meshheading:8995312-Saccharomyces cerevisiae, pubmed-meshheading:8995312-Saccharomyces cerevisiae Proteins, pubmed-meshheading:8995312-Sequence Alignment, pubmed-meshheading:8995312-Serine, pubmed-meshheading:8995312-Structure-Activity Relationship, pubmed-meshheading:8995312-Substrate Specificity, pubmed-meshheading:8995312-Transferases, pubmed-meshheading:8995312-Tyrosine
pubmed:year
1997
pubmed:articleTitle
Amino acid substitutions that convert the protein substrate specificity of farnesyltransferase to that of geranylgeranyltransferase type I.
pubmed:affiliation
Department of Microbiology and Molecular Genetics, Molecular Biology Institute, UCLA, Los Angeles, California 90095-1489, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.