Cloning and expression of Escherichia coli 5'-methylthioadenosine/S- adenosylhomocysteine nucleosidase identification of the pfs gene product

Kenneth A. Cornell; Michael K. Riscoe

doi:10.1016/S0167-4781(97)00169-3

Cloning and expression of Escherichia coli 5'-methylthioadenosine/S- adenosylhomocysteine nucleosidase identification of the pfs gene product

Kenneth A. Cornell, Michael K. Riscoe

Chemistry Department

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

46 Scopus citations

Abstract

The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (EC 3.2.2.9) is responsible for cleavage of the glycosidic bond in both 5'- methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH). Based on amino acid sequence analysis of this enzyme from Klebsiella, we recently speculated that an open reading frame found in E. coli (designated pfs) encoded MTA/SAH nucleosidase. To explore this possibility, we amplified, cloned, and expressed the complete pfs gene from E. coli genomic DNA. The recombinant protein exhibited a molecular weight and Michaelis constants for MTA that are in agreement with those reported for native enzyme. From this biochemical evidence we confirm our original assignment of the pfs gene as encoding MTA/SAH nucleosidase.

Original language	English
Pages (from-to)	8-14
Number of pages	7
Journal	Biochimica et Biophysica Acta - Gene Structure and Expression
Volume	1396
Issue number	1
DOIs	https://doi.org/10.1016/S0167-4781(97)00169-3
State	Published - 4 Mar 1998

Keywords

(E. coli)
Methionine
MTA phosphorylase
MTA/SAH nucleosidase
MTR kinase
Pfs gene
Salvage

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10.1016/S0167-4781(97)00169-3

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title = "Cloning and expression of Escherichia coli 5'-methylthioadenosine/S- adenosylhomocysteine nucleosidase identification of the pfs gene product",

abstract = "The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (EC 3.2.2.9) is responsible for cleavage of the glycosidic bond in both 5'- methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH). Based on amino acid sequence analysis of this enzyme from Klebsiella, we recently speculated that an open reading frame found in E. coli (designated pfs) encoded MTA/SAH nucleosidase. To explore this possibility, we amplified, cloned, and expressed the complete pfs gene from E. coli genomic DNA. The recombinant protein exhibited a molecular weight and Michaelis constants for MTA that are in agreement with those reported for native enzyme. From this biochemical evidence we confirm our original assignment of the pfs gene as encoding MTA/SAH nucleosidase.",

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T1 - Cloning and expression of Escherichia coli 5'-methylthioadenosine/S- adenosylhomocysteine nucleosidase identification of the pfs gene product

AU - Cornell, Kenneth A.

AU - Riscoe, Michael K.

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N2 - The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (EC 3.2.2.9) is responsible for cleavage of the glycosidic bond in both 5'- methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH). Based on amino acid sequence analysis of this enzyme from Klebsiella, we recently speculated that an open reading frame found in E. coli (designated pfs) encoded MTA/SAH nucleosidase. To explore this possibility, we amplified, cloned, and expressed the complete pfs gene from E. coli genomic DNA. The recombinant protein exhibited a molecular weight and Michaelis constants for MTA that are in agreement with those reported for native enzyme. From this biochemical evidence we confirm our original assignment of the pfs gene as encoding MTA/SAH nucleosidase.

AB - The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (EC 3.2.2.9) is responsible for cleavage of the glycosidic bond in both 5'- methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH). Based on amino acid sequence analysis of this enzyme from Klebsiella, we recently speculated that an open reading frame found in E. coli (designated pfs) encoded MTA/SAH nucleosidase. To explore this possibility, we amplified, cloned, and expressed the complete pfs gene from E. coli genomic DNA. The recombinant protein exhibited a molecular weight and Michaelis constants for MTA that are in agreement with those reported for native enzyme. From this biochemical evidence we confirm our original assignment of the pfs gene as encoding MTA/SAH nucleosidase.

KW - (E. coli)

KW - Methionine

KW - MTA phosphorylase

KW - MTA/SAH nucleosidase

KW - MTR kinase

KW - Pfs gene

KW - Salvage

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U2 - 10.1016/S0167-4781(97)00169-3

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JO - Biochimica et Biophysica Acta - Gene Structure and Expression

JF - Biochimica et Biophysica Acta - Gene Structure and Expression

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ER -

Cloning and expression of Escherichia coli 5'-methylthioadenosine/S- adenosylhomocysteine nucleosidase identification of the pfs gene product

Abstract

Keywords

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