Synthesis and enzymatic studies of bisubstrate analogues for farnesyl diphosphate synthase.
Farnesyl diphosphate synthase catalyzes the sequential chain elongation reactions between isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) to form geranyl diphosphate (GPP) and between IPP and GPP to give farnesyl diphosphate (FPP). Bisubstrate analogues containing the allylic and homoallylic substrates were synthesized by joining fragments for IPP and the allylic diphosphates with a C-C bond between the methyl group at C3 in IPP and the Z-methyl group at C3 in DMAPP (3-OPP) and GPP (4-OPP), respectively. These constructs placed substantial limits on the conformational space available to the analogues relative to the two substrates. The key features of the synthesis of bisubstrate analogues 3-OPP and 4-OPP are a regioselective C-alkylation of the dianion of 3-methyl-3-buten-1-ol (5), a Z-selective cuprate addition of alkyl groups to an α,β-alkynyl ester intermediate, and differential activation of allylic and homoallylic alcohols in the analogues, followed by a simultaneous displacement of the leaving groups with tris(tetra-n-butylammonium) hydrogen diphosphate to give the corresponding bisdiphosphate analogues. The bisubstrate analogues were substrates for FPP synthase, giving novel seven-membered ring analogues of GPP and FPP. The catalytic efficiencies for cyclization of 3-OPP and 4-OPP were similar to those for chain elongation with IPP and DMAPP.
†Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
Gurusankar
G
Ramamoorthy
†Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
Mark L
ML
Pugh
‡Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, California 94158, United States.|§California Institute for Quantitative Biomedical Research, University of California, San Francisco, California 94158, United States.
Bo-Xue
BX
Tian
†Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
Richard M
RM
Phan
†Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
Lawrence B
LB
Perez
‡Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, California 94158, United States.|§California Institute for Quantitative Biomedical Research, University of California, San Francisco, California 94158, United States.
Matthew P
MP
Jacobson
†Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
C Dale
CD
Poulter
D000440:Butanols / Q000737:chemistry*; D002384:Catalysis; D003500:Cyclization; D051228:Geranyltranstransferase / Q000138:chemical synthesis* / Q000737:chemistry; D011106:Polyisoprenyl Phosphates / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012717:Sesquiterpenes / Q000737:chemistry*; D013379:Substrate Specificity
D000440:Butanols; D011106:Polyisoprenyl Phosphates; D000644:Quaternary Ammonium Compounds; D012717:Sesquiterpenes; C000603534:tris(tetra-n-butylammonium)hydrogen diphosphate; C004808:farnesyl pyrophosphate; D051228:Geranyltranstransferase; C000603533:3-methyl-3-buten-1-ol