Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells

Hyo I. Chang; Nadine D. Cohen

doi:10.1016/0003-9861(83)90026-7

Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells

Hyo I. Chang, Nadine D. Cohen

Research output: Contribution to journal › Article › peer-review

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Abstract

A quantitative assay has been developed to measure holocarboxylase synthetase activity in cellular extracts. This assay was based on measuring the incorporation of [³H]biotin of high specific activity (4.3 Ci/mmol) into purified rat liver apopyruvate carboxylase. With this assay, holocarboxylase synthetase in 3T3-L1 mouse fibroblasts has been monitored. During the differentiation of this cell from a fibroblast to an adipocyte, holocarboxylase synthetase activity was found to increase threefold, while pyruvate carboxylase activity rose 20-fold. The results suggest a possible relationship between the activity of the holocarboxylase synthetase and the level of the biotin-dependent carboxylases within the mammalian cell. Utilizing digitonin fractionation, the intracellular distribution of this enzyme has also been examined. In the 3T3-L1 cell, the large majority (approximately 70%) of the total holocarboxylase synthetase activity was found in the cytosolic compartment.

Original language	English
Pages (from-to)	237-247
Number of pages	11
Journal	Archives of Biochemistry and Biophysics
Volume	225
Issue number	1
DOIs	https://doi.org/10.1016/0003-9861(83)90026-7
Publication status	Published - 1983 Aug
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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@article{f37e404f3250452c9726885559482b6a,

title = "Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells",

abstract = "A quantitative assay has been developed to measure holocarboxylase synthetase activity in cellular extracts. This assay was based on measuring the incorporation of [3H]biotin of high specific activity (4.3 Ci/mmol) into purified rat liver apopyruvate carboxylase. With this assay, holocarboxylase synthetase in 3T3-L1 mouse fibroblasts has been monitored. During the differentiation of this cell from a fibroblast to an adipocyte, holocarboxylase synthetase activity was found to increase threefold, while pyruvate carboxylase activity rose 20-fold. The results suggest a possible relationship between the activity of the holocarboxylase synthetase and the level of the biotin-dependent carboxylases within the mammalian cell. Utilizing digitonin fractionation, the intracellular distribution of this enzyme has also been examined. In the 3T3-L1 cell, the large majority (approximately 70%) of the total holocarboxylase synthetase activity was found in the cytosolic compartment.",

author = "Chang, {Hyo I.} and Cohen, {Nadine D.}",

note = "Funding Information: i These studies were supported in part by research grants from the National Institutes of Health (AM 38651) and National Science Foundation (PCM 8103270). {\textquoteleft}To whom requests for reprints should be addressed.",

year = "1983",

month = aug,

doi = "10.1016/0003-9861(83)90026-7",

language = "English",

volume = "225",

pages = "237--247",

journal = "Archives of Biochemistry and Biophysics",

issn = "0003-9861",

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T1 - Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells

AU - Chang, Hyo I.

AU - Cohen, Nadine D.

N1 - Funding Information: i These studies were supported in part by research grants from the National Institutes of Health (AM 38651) and National Science Foundation (PCM 8103270). ‘To whom requests for reprints should be addressed.

PY - 1983/8

Y1 - 1983/8

N2 - A quantitative assay has been developed to measure holocarboxylase synthetase activity in cellular extracts. This assay was based on measuring the incorporation of [3H]biotin of high specific activity (4.3 Ci/mmol) into purified rat liver apopyruvate carboxylase. With this assay, holocarboxylase synthetase in 3T3-L1 mouse fibroblasts has been monitored. During the differentiation of this cell from a fibroblast to an adipocyte, holocarboxylase synthetase activity was found to increase threefold, while pyruvate carboxylase activity rose 20-fold. The results suggest a possible relationship between the activity of the holocarboxylase synthetase and the level of the biotin-dependent carboxylases within the mammalian cell. Utilizing digitonin fractionation, the intracellular distribution of this enzyme has also been examined. In the 3T3-L1 cell, the large majority (approximately 70%) of the total holocarboxylase synthetase activity was found in the cytosolic compartment.

AB - A quantitative assay has been developed to measure holocarboxylase synthetase activity in cellular extracts. This assay was based on measuring the incorporation of [3H]biotin of high specific activity (4.3 Ci/mmol) into purified rat liver apopyruvate carboxylase. With this assay, holocarboxylase synthetase in 3T3-L1 mouse fibroblasts has been monitored. During the differentiation of this cell from a fibroblast to an adipocyte, holocarboxylase synthetase activity was found to increase threefold, while pyruvate carboxylase activity rose 20-fold. The results suggest a possible relationship between the activity of the holocarboxylase synthetase and the level of the biotin-dependent carboxylases within the mammalian cell. Utilizing digitonin fractionation, the intracellular distribution of this enzyme has also been examined. In the 3T3-L1 cell, the large majority (approximately 70%) of the total holocarboxylase synthetase activity was found in the cytosolic compartment.

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DO - 10.1016/0003-9861(83)90026-7

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JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 1

ER -

Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3-L1 cells

Abstract

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