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Molecular cloning of CoA synthase - the missing link in CoA biosynthesis

Molecular cloning of CoA synthase - the missing link in CoA biosynthesis
Molecular cloning of CoA synthase - the missing link in CoA biosynthesis
Coenzyme A functions as a carrier of acetyl and acyl groups in living cells and is essential for numerous biosynthetic, energy-yielding, and degradative metabolic pathways. There are five enzymatic steps in CoA biosynthesis. To date, molecular cloning of enzymes involved in the CoA biosynthetic pathway in mammals has been only reported for pantothenate kinase. In this study, we present cDNA cloning and functional characterization of CoA synthase. It has an open reading frame of 563 aa and encodes a protein of similar to60 kDa. Sequence alignments suggested that the protein possesses both phosphopantetheine adenylyltransferase and dephospho-CoA kinase domains. Biochemical assays using wild type recombinant protein confirmed the gene product indeed contained both these enzymatic activities. The presence of intrinsic phosphopantetheine adenylyltransferase activity was further confirmed by site-directed mutagenesis. Therefore, this study describes the first cloning and characterization of a mammalian CoA synthase and confirms this is a bifunctional enzyme containing the last two components of CoA biosynthesis.
PANTOTHENATE KINASE GENE, COENZYME-A METABOLISM, ESCHERICHIA-COLI, RAT-LIVER, ADENYLYLTRANSFERASE, HEART, PURIFICATION, COMPLEX, MUSCLE, STATE
1083-351X
22107 - 22110
Zhyvoloup, A
5b0b4802-af34-4399-94e7-da217bda2538
Nemazanyy, I
b0613ad8-2293-4a71-9b3d-d700f9366de4
Babich, A
81b29b74-5751-4fdc-8a2f-475b3e50be8e
Panasyuk, G
b6601a2f-3f81-4cb1-9c87-fe8b26d007e0
Pobigailo, N
d6f83d68-f4b9-4c8f-b239-5720efe9a12d
Vudmaska, M
ccb9a21a-0fdc-4e60-b852-bbef606ac587
Naidenov, V
dd48d604-e23d-4f9f-b988-78dc5e4889b4
Kukharenko, O
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Palchevskii, S
a5a993fa-20a0-4ebc-b998-227dd8d1eb5d
Savinska, L
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Ovcharenko, G
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Verdier, F
f07a780c-0dc5-4180-ab56-3c860b5f80ab
Valovka, T
97346de1-eac0-4e81-b82e-b39f835474f7
Fenton, TR
087260ba-f6a1-405a-85df-099d05810a84
Rebholz, H
1dcf329e-8cfd-4034-a28d-ec2b2c3c946f
Wang, ML
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Shepherd, P
5dbcbb99-0e1a-464d-b24f-91a1dfea735a
Matsuka, G
5c9c20b5-446f-4a5c-b2b3-20219d9cd5d5
Filonenko, V
19bb8baa-42b2-41a3-87bc-0cdcccf2f9b6
Gout, IT
7fd0e0ff-a6a5-4579-aea1-fa3c1e80fe23
Zhyvoloup, A
5b0b4802-af34-4399-94e7-da217bda2538
Nemazanyy, I
b0613ad8-2293-4a71-9b3d-d700f9366de4
Babich, A
81b29b74-5751-4fdc-8a2f-475b3e50be8e
Panasyuk, G
b6601a2f-3f81-4cb1-9c87-fe8b26d007e0
Pobigailo, N
d6f83d68-f4b9-4c8f-b239-5720efe9a12d
Vudmaska, M
ccb9a21a-0fdc-4e60-b852-bbef606ac587
Naidenov, V
dd48d604-e23d-4f9f-b988-78dc5e4889b4
Kukharenko, O
8e63ef16-3394-4072-96f9-953283db0b6f
Palchevskii, S
a5a993fa-20a0-4ebc-b998-227dd8d1eb5d
Savinska, L
daa1bea5-62d1-4749-a99a-13fa6bfd6e49
Ovcharenko, G
56af406a-dcb9-4c72-a44c-04186395c895
Verdier, F
f07a780c-0dc5-4180-ab56-3c860b5f80ab
Valovka, T
97346de1-eac0-4e81-b82e-b39f835474f7
Fenton, TR
087260ba-f6a1-405a-85df-099d05810a84
Rebholz, H
1dcf329e-8cfd-4034-a28d-ec2b2c3c946f
Wang, ML
94be8ea1-1dd1-4d64-a01c-86209c94e5d1
Shepherd, P
5dbcbb99-0e1a-464d-b24f-91a1dfea735a
Matsuka, G
5c9c20b5-446f-4a5c-b2b3-20219d9cd5d5
Filonenko, V
19bb8baa-42b2-41a3-87bc-0cdcccf2f9b6
Gout, IT
7fd0e0ff-a6a5-4579-aea1-fa3c1e80fe23

Zhyvoloup, A, Nemazanyy, I, Babich, A, Panasyuk, G, Pobigailo, N, Vudmaska, M, Naidenov, V, Kukharenko, O, Palchevskii, S, Savinska, L, Ovcharenko, G, Verdier, F, Valovka, T, Fenton, TR, Rebholz, H, Wang, ML, Shepherd, P, Matsuka, G, Filonenko, V and Gout, IT (2002) Molecular cloning of CoA synthase - the missing link in CoA biosynthesis. Journal of Biological Chemistry, 277 (25), 22107 - 22110. (doi:10.1074/jbc.C200195200).

Record type: Article

Abstract

Coenzyme A functions as a carrier of acetyl and acyl groups in living cells and is essential for numerous biosynthetic, energy-yielding, and degradative metabolic pathways. There are five enzymatic steps in CoA biosynthesis. To date, molecular cloning of enzymes involved in the CoA biosynthetic pathway in mammals has been only reported for pantothenate kinase. In this study, we present cDNA cloning and functional characterization of CoA synthase. It has an open reading frame of 563 aa and encodes a protein of similar to60 kDa. Sequence alignments suggested that the protein possesses both phosphopantetheine adenylyltransferase and dephospho-CoA kinase domains. Biochemical assays using wild type recombinant protein confirmed the gene product indeed contained both these enzymatic activities. The presence of intrinsic phosphopantetheine adenylyltransferase activity was further confirmed by site-directed mutagenesis. Therefore, this study describes the first cloning and characterization of a mammalian CoA synthase and confirms this is a bifunctional enzyme containing the last two components of CoA biosynthesis.

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More information

e-pub ahead of print date: 29 April 2002
Published date: 21 June 2002
Keywords: PANTOTHENATE KINASE GENE, COENZYME-A METABOLISM, ESCHERICHIA-COLI, RAT-LIVER, ADENYLYLTRANSFERASE, HEART, PURIFICATION, COMPLEX, MUSCLE, STATE

Identifiers

Local EPrints ID: 453962
URI: http://eprints.soton.ac.uk/id/eprint/453962
ISSN: 1083-351X
PURE UUID: 6bcbb357-3afb-4e89-8c08-d43204fc1245
ORCID for TR Fenton: ORCID iD orcid.org/0000-0002-4737-8233

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Date deposited: 26 Jan 2022 17:50
Last modified: 17 Mar 2024 04:11

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Contributors

Author: A Zhyvoloup
Author: I Nemazanyy
Author: A Babich
Author: G Panasyuk
Author: N Pobigailo
Author: M Vudmaska
Author: V Naidenov
Author: O Kukharenko
Author: S Palchevskii
Author: L Savinska
Author: G Ovcharenko
Author: F Verdier
Author: T Valovka
Author: TR Fenton ORCID iD
Author: H Rebholz
Author: ML Wang
Author: P Shepherd
Author: G Matsuka
Author: V Filonenko
Author: IT Gout

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