Analysis of isoprenoid biosynthesis in peroxisomal-deficient Pex2 CHO cell lines.

Abstract:

:ZR-78 and ZR-82 cells are two peroxisomal-deficient Chinese hamster ovary (CHO) cell mutants. These cells lack normal peroxisomes and show reduced levels of plasmalogen synthesis and other peroxisomal functions attributed to the deficiency of peroxisomal matrix enzymes. As we have recently identified two HMG-CoA reductase proteins in CHO cells, a 97 kDa reductase localized in the ER and a 90 kDa reductase protein localized in peroxisomes, this enabled us to study the two reductase proteins for the first time in peroxisomal-deficient CHO cells. In this study we report the results of a detailed analysis of the isoprenoid biosynthetic pathway in the peroxisomal-deficient CHO cell lines ZR-78 and ZR-82. We demonstrate that total HMG-CoA reductase activity is significantly reduced in the peroxisomal-deficient cells as compared to the wild type cells. Analysis of the two reductase proteins in permeabilized cells indicated that in the ZR-78 and ZR-82 cells the 90 kDa peroxisomal reductase protein was mainly localized to the cytosol. We further show that the rates of both sterol (cholesterol) and non-sterol (dolichols) biosynthesis were significantly lower in the peroxisomal-deficient cells, when either [3H] acetate or [3H] mevalonate was used as substrate. In contrast, the rate of dolichol biosynthesis in the peroxisomal-deficient cells was similar to that of the wild type cells when incubated with [3H] farnesol. In addition, we demonstrate that the peroxisomal-deficient cells exhibited increased rates of lanosterol biosynthesis as compared to wild type cells. The results of this study provide further evidence for the essential requirement of peroxisomes for cholesterol biosynthesis as well as for dolichol production.

journal_name

J Lipid Res

authors

Aboushadi N,Krisans SK

subject

Has Abstract

pub_date

1998-09-01 00:00:00

pages

1781-91

issue

9

eissn

0022-2275

issn

1539-7262

journal_volume

39

pub_type

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