Document Detail


A role of the amino acid residue located on the fifth position before the first aspartate-rich motif of farnesyl diphosphate synthase on determination of the final product.
MedLine Citation:
PMID:  8940054     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Farnesyl diphosphate (FPP) synthase catalyzes consecutive condensations of isopentenyl diphosphate with allylic substrates to give FPP, C-15 compound, as a final product and does not catalyze a condensation beyond FPP. Recently, it was observed that, in Bacillus stearothermophilus FPP synthase, a replacement of tyrosine with histidine at position 81, which is located on the fifth amino acid before the first aspartate-rich motif, caused the mutated FPP synthase to catalyze geranylgeranyl diphosphate (C-20) synthesis (Ohnuma, S.-i., Nakazawa, T., Hemmi, H., Hallberg, A.-M., Koyama, T., Ogura, K., and Nishino, T. (1996) J. Biol. Chem. 271, 10087-10095). Thus, we constructed 20 FPP synthases, each of which has a different amino acid at position 81, and analyzed them. All enzymes except for Y81P can catalyze the condensations of isopentenyl diphosphate. The final products and the product distributions are different from each other. Y81A, Y81G, and Y81S can produce hexaprenyl diphosphate (C-30) as their final product. The final product of Y81C, Y81H, Y81I, Y81L, Y81N, Y81T, and Y81V are geranylfarnesyl diphosphate (C-25), and Y81D, Y81E, Y81F, Y81K, Y81M, Y81Q, and Y81R cannot produce polyprenyl diphosphates more than geranylgeranyl diphosphate. Substitution of tryptophan does not affect the product specificity of FPP synthase. The average chain length of products is inversely proportional to the accessible surface area of substituted amino acid. However, no significant relation between the final chain length and the kinetic constants Km and Vmax are observed. These observations strongly indicate that the amino acid does not come into contact with the substrates but directly contacts the omega-terminal of an elongating allylic product. This interaction must prevent further condensation of isopentenyl diphosphate.
Authors:
S i Ohnuma; K Narita; T Nakazawa; C Ishida; Y Takeuchi; C Ohto; T Nishino
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  271     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1996 Nov 
Date Detail:
Created Date:  1997-01-07     Completed Date:  1997-01-07     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  30748-54     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Engineering, Tohoku University, Aoba Aramaki, Aoba-ku, Sendai 980-77 Japan. sohnuma@seika.che.tohoku.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Alkyl and Aryl Transferases*
Amino Acid Sequence
Binding Sites
Geobacillus stearothermophilus / enzymology*
Geranyltranstransferase
Kinetics
Mutagenesis, Site-Directed
Polyisoprenyl Phosphates / metabolism
Structure-Activity Relationship
Substrate Specificity
Surface Properties
Transferases / chemistry*,  metabolism
Chemical
Reg. No./Substance:
0/Polyisoprenyl Phosphates; EC 2.-/Transferases; EC 2.5.-/Alkyl and Aryl Transferases; EC 2.5.1.10/Geranyltranstransferase

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