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

Molecular Microbiology and Immunology

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

45 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 (US)
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 - Mar 4 1998

Keywords

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

ASJC Scopus subject areas

Structural Biology
Biophysics
Biochemistry
Genetics

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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.",

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

author = "Cornell, {Kenneth A.} and Riscoe, {Michael K.}",

note = "Funding Information: We gratefully acknowledge support from the Veterans' Affairs Medical Research Program and financial contributions by the Collins Medical Trust of Oregon, the Medical Research Foundation of Oregon, and Interlab (Lake Oswego, OR). KC receives support from the National Institute of Health Molecular Hematology Training Program Grant #T32-HL07781 awarded to the Oregon Health Sciences University. ",

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doi = "10.1016/S0167-4781(97)00169-3",

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AU - Riscoe, Michael K.

N1 - Funding Information: We gratefully acknowledge support from the Veterans' Affairs Medical Research Program and financial contributions by the Collins Medical Trust of Oregon, the Medical Research Foundation of Oregon, and Interlab (Lake Oswego, OR). KC receives support from the National Institute of Health Molecular Hematology Training Program Grant #T32-HL07781 awarded to the Oregon Health Sciences University.

PY - 1998/3/4

Y1 - 1998/3/4

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.

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KW - MTR kinase

KW - Methionine

KW - Pfs gene

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

Abstract

Keywords

ASJC Scopus subject areas

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