PMID- 20730526
OWN - NLM
STAT- MEDLINE
DCOM- 20110105
LR  - 20191210
IS  - 1618-2650 (Electronic)
IS  - 1618-2642 (Linking)
VI  - 398
IP  - 4
DP  - 2010 Oct
TI  - A rapid and sensitive assay for determining human brain levels of farnesyl-(FPP) 
      and geranylgeranylpyrophosphate (GGPP) and transferase activities using 
      UHPLC-MS/MS.
PG  - 1801-8
LID - 10.1007/s00216-010-4088-7 [doi]
AB  - The isoprenoids farnesyl-(FPP) and geranylgeranylpyrophosphate (FPP and GGPP) are 
      two major lipid intermediates in the mevalonate pathway. They participate in 
      post-translational modification of members of the superfamily of small guanosine 
      triphosphatases (GTPases; Ras, Rab, Rac, etc.) via prenylation reactions. Due to 
      the important role of these proteins in a number of cell processes, in particular 
      cell growth, division, and differentiation, investigation of the involvement of 
      isoprenoids in these processes is of great interest. In a previously published 
      report, we described a fully validated assay for the quantitation of the two 
      isoprenoids using a high-performance liquid chromatography (HPLC)-fluorescence 
      detection (FLD) method. The current work expands on the previous method and 
      enhances it greatly by using a much faster state-of-the-art ultrahigh-performance 
      liquid chromatography (UHPLC) technique coupled to tandem mass spectrometry 
      (MS/MS). The method exhibited a linear concentration range of 5-250 ng/mL for FPP 
      and GGPP in human brain tissue; it was shown to be unaffected by ion suppression 
      and provided results almost six times faster than the HPLC-FLD assay. Comparison 
      of UHPLC-MS/MS and HPLC-FLD yielded excellent comparability of the two assays for 
      both isoprenoids. Based on the UHPLC-MS/MS assay, a novel in vitro test system 
      was implemented to study enzyme specificity for distinct amino acid CAAX motifs, 
      which is potentially useful for investigating target interactions of new 
      therapeutics for diseases involving pathological regulation of isoprenoids and/or 
      small GTPases.
FAU - Hooff, Gero P
AU  - Hooff GP
AD  - Department of Pharmacology, Biocenter Niederursel, University of Frankfurt, 
      Max-von Laue-Str. 9, 60438 Frankfurt, Germany.
FAU - Patel, Nina
AU  - Patel N
FAU - Wood, W Gibson
AU  - Wood WG
FAU - Muller, Walter E
AU  - Muller WE
FAU - Eckert, Gunter P
AU  - Eckert GP
FAU - Volmer, Dietrich A
AU  - Volmer DA
LA  - eng
GR  - AG18357/AG/NIA NIH HHS/United States
GR  - AG23524/AG/NIA NIH HHS/United States
GR  - Medical Research Council/United Kingdom
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20100821
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Polyisoprenyl Phosphates)
RN  - 0 (Sesquiterpenes)
RN  - 79W6B01D07 (farnesyl pyrophosphate)
RN  - EC 2.5.- (Alkyl and Aryl Transferases)
RN  - EC 2.5.1.- (geranylgeranyltransferase type-I)
RN  - EC 2.5.1.21 (Farnesyl-Diphosphate Farnesyltransferase)
RN  - N21T0D88LX (geranylgeranyl pyrophosphate)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Alkyl and Aryl Transferases/*chemistry/metabolism
MH  - Brain/enzymology/metabolism
MH  - *Brain Chemistry
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Farnesyl-Diphosphate Farnesyltransferase/*chemistry/metabolism
MH  - Female
MH  - Humans
MH  - Kinetics
MH  - Male
MH  - Middle Aged
MH  - Polyisoprenyl Phosphates/*analysis/metabolism
MH  - Sesquiterpenes/*analysis/metabolism
MH  - Tandem Mass Spectrometry/*methods
EDAT- 2010/08/24 06:00
MHDA- 2011/01/06 06:00
CRDT- 2010/08/24 06:00
PHST- 2010/07/08 00:00 [received]
PHST- 2010/08/02 00:00 [accepted]
PHST- 2010/07/30 00:00 [revised]
PHST- 2010/08/24 06:00 [entrez]
PHST- 2010/08/24 06:00 [pubmed]
PHST- 2011/01/06 06:00 [medline]
AID - 10.1007/s00216-010-4088-7 [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2010 Oct;398(4):1801-8. doi: 10.1007/s00216-010-4088-7. Epub 
      2010 Aug 21.