Quantitative determination of farnesyl and geranylgeranyl diphosphate levels in mammalian tissue

Huaxiang Tong, Andrew J. Wiemer, Jeffrey Neighbors, Raymond Hohl

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) are branch point intermediates of isoprenoid biosynthesis. Inhibitors of isoprenoid biosynthesis, such as the statins and bisphosphonates, are widely used therapeutic agents. However, little is known about the degree to which they alter levels of upstream and downstream isoprenoids, including FPP and GGPP. Therefore, we developed a method to isolate and quantify FPP and GGPP from mammalian tissues. Tissues from mice were collected, snap frozen in liquid nitrogen, and stored at -80 °C. FPP and GGPP were isolated by a combined homogenization and extraction procedure and were purified with a C18 solid phase extraction column. Farnesyl protein transferase (FTase) or geranylgeranyl protein transferase I (GGTase I) were used to conjugate FPP and GGPP with fluorescent dansylated peptides. FPP and GGPP were quantified by high-performance liquid chromatography (HPLC). The respective concentrations of FPP and GGPP are as follows: 0.355 ± 0.030 and 0.827 ± 0.082 units of nmol/g wet tissues in brain, 0.320 ± 0.019 and 0.293 ± 0.035 units of nmol/g wet tissues in kidney, 0.326 ± 0.064 and 0.213 ± 0.029 units of nmol/g wet tissues in liver, and 0.364 ± 0.015 and 0.349 ± 0.023 units of nmol/g wet tissues in heart (means ± SEM). This method allows for determination of FPP and GGPP concentrations in any tissue type and is sensitive enough to detect changes following treatment with inhibitors of isoprenoid biosynthesis.

Original languageEnglish (US)
Pages (from-to)138-143
Number of pages6
JournalAnalytical Biochemistry
Volume378
Issue number2
DOIs
StatePublished - Jul 15 2008

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Tissue
Terpenes
Biosynthesis
Transferases
Hydroxymethylglutaryl-CoA Reductase Inhibitors
farnesyl pyrophosphate
geranylgeranyl pyrophosphate
Solid Phase Extraction
Diphosphonates
High performance liquid chromatography
Liquid nitrogen
Liver
Brain
Proteins
Nitrogen
High Pressure Liquid Chromatography
Kidney
Peptides
Scanning electron microscopy

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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title = "Quantitative determination of farnesyl and geranylgeranyl diphosphate levels in mammalian tissue",
abstract = "Farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) are branch point intermediates of isoprenoid biosynthesis. Inhibitors of isoprenoid biosynthesis, such as the statins and bisphosphonates, are widely used therapeutic agents. However, little is known about the degree to which they alter levels of upstream and downstream isoprenoids, including FPP and GGPP. Therefore, we developed a method to isolate and quantify FPP and GGPP from mammalian tissues. Tissues from mice were collected, snap frozen in liquid nitrogen, and stored at -80 °C. FPP and GGPP were isolated by a combined homogenization and extraction procedure and were purified with a C18 solid phase extraction column. Farnesyl protein transferase (FTase) or geranylgeranyl protein transferase I (GGTase I) were used to conjugate FPP and GGPP with fluorescent dansylated peptides. FPP and GGPP were quantified by high-performance liquid chromatography (HPLC). The respective concentrations of FPP and GGPP are as follows: 0.355 ± 0.030 and 0.827 ± 0.082 units of nmol/g wet tissues in brain, 0.320 ± 0.019 and 0.293 ± 0.035 units of nmol/g wet tissues in kidney, 0.326 ± 0.064 and 0.213 ± 0.029 units of nmol/g wet tissues in liver, and 0.364 ± 0.015 and 0.349 ± 0.023 units of nmol/g wet tissues in heart (means ± SEM). This method allows for determination of FPP and GGPP concentrations in any tissue type and is sensitive enough to detect changes following treatment with inhibitors of isoprenoid biosynthesis.",
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Quantitative determination of farnesyl and geranylgeranyl diphosphate levels in mammalian tissue. / Tong, Huaxiang; Wiemer, Andrew J.; Neighbors, Jeffrey; Hohl, Raymond.

In: Analytical Biochemistry, Vol. 378, No. 2, 15.07.2008, p. 138-143.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Quantitative determination of farnesyl and geranylgeranyl diphosphate levels in mammalian tissue

AU - Tong, Huaxiang

AU - Wiemer, Andrew J.

AU - Neighbors, Jeffrey

AU - Hohl, Raymond

PY - 2008/7/15

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