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Histochemistry and Cell Biology

, Volume 127, Issue 3, pp 273–290 | Cite as

Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

  • Werner J. Kovacs
  • Khanichi N. Tape
  • Janis E. Shackelford
  • Xueying Duan
  • Takhar Kasumov
  • Joanne K. Kelleher
  • Henri Brunengraber
  • Skaidrite K. Krisans
Original Paper
First Online:
Accepted:

Abstract

Previous studies have indicated that the early steps in the isoprenoid/cholesterol biosynthetic pathway occur in peroxisomes. However, the role of peroxisomes in cholesterol biosynthesis has recently been questioned in several reports concluding that three of the peroxisomal cholesterol biosynthetic enzymes, namely mevalonate kinase, phosphomevalonate kinase, and mevalonate diphosphate decarboxylase, do not localize to peroxisomes in human cells even though they contain consensus peroxisomal targeting signals. We re-investigated the subcellular localization of the cholesterol biosynthetic enzymes of the pre-squalene segment in human cells by using new stable isotopic techniques and data computations with isotopomer spectral analysis, in combination with immunofluorescence and cell permeabilization techniques. Our present findings clearly show and confirm previous studies that the pre-squalene segment of the cholesterol biosynthetic pathway is localized to peroxisomes. In addition, our data are consistent with the hypothesis that acetyl-CoA derived from peroxisomal β-oxidation of very long-chain fatty acids and medium-chain dicarboxylic acids is preferentially channeled to cholesterol synthesis inside the peroxisomes without mixing with the cytosolic acetyl-CoA pool.

Keywords

Cholesterol biosynthesis Isoprenoid Peroxisomes Mass isotopomer distribution HepG2 

Abbreviations

AOX

Acyl-CoA oxidase

FPP

Farnesyl diphosphate

GFP

Green fluorescent protein

HMGCR

HMG-CoA reductase

IDI1

Isopentenyl diphosphate isomerase 1

ISA

Isotopomer spectral analysis

MID

Mass isotopomer distributions

MPD

Mevalonate diphosphate decarboxylase

MVK

Mevalonate kinase

PEX

Peroxin

PMP

Peroxisomal membrane protein

PMVK

Phosphomevalonate kinase

PP

Diphosphate

PTS

Peroxisomal targeting signal

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Notes

Acknowledgments

This work was supported by National Institutes of Health (NIH) grants DK58238 and DK58040 to S.K.K. We thank Drs. Eveline Baumgart-Vogt and Herbert Stangl for helpful discussions and comments on the manuscript.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Werner J. Kovacs
    • 1
    • 2
    • 5
  • Khanichi N. Tape
    • 1
  • Janis E. Shackelford
    • 1
  • Xueying Duan
    • 3
  • Takhar Kasumov
    • 3
  • Joanne K. Kelleher
    • 4
  • Henri Brunengraber
    • 3
  • Skaidrite K. Krisans
    • 1
  1. 1.Department of BiologySan Diego State UniversitySan DiegoUSA
  2. 2.Department of Anatomy and Cell Biology, Division of Medical Cell BiologyJustus Liebig University of GiessenGiessenGermany
  3. 3.Department of NutritionCase Western Reserve UniversityClevelandUSA
  4. 4.Department of Biomedical EngineeringMassachusetts Institute of TechnologyCambridgeUSA
  5. 5.Institute of Cell BiologyZürichSwitzerland

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