Characterization of IMT1, myo-Inositol O-methyltransferase, from Mesembryanthemum crystallinum
A full-length transcript, Imt1, encoding myo-inositol O-methyltransferase (EC 2.1.1.X) from the halophyte Mesembryanthemum crystallinum was expressed in Escherichia coli. The enzyme, IMT1, uses S-adenosyl-L-methionine to methylate myo-inositol to form D-ononitol. IMT1 with a monomeric mass of 41,000...
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| Veröffentlicht in: | Archives of biochemistry and biophysics 1995-09, Vol.322 (1), p.183-188 |
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| creator | Rammesmayer, G. Pichorner, H. Adams, P. Jensen, R.G. Bohnert, H.J. |
| description | A full-length transcript,
Imt1, encoding
myo-inositol
O-methyltransferase (EC 2.1.1.X) from the halophyte
Mesembryanthemum crystallinum was expressed in
Escherichia coli. The enzyme, IMT1, uses
S-adenosyl-L-methionine to methylate
myo-inositol to form D-ononitol. IMT1 with a monomeric mass of 41,000 was isolated by ammonium sulfate fractionation, gel filtration and ion exchange chromatography to apparent purity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified recombinant enzyme was identical to that encoded by the cDNA sequence. The apparent
K
m
for
S-adenosylmethionine was 0.18 mM with a
V
max of 1550 pkat/mg protein. The
K
m
for
myo-inositol was 1.32 mM. The reaction became substrate-inhibited by concentrations of
S-adenosylmethionine greater than 0.5 mM. Inositol methyltransferase was competitively inhibited 50% with 0.01 mM
S-adenosyl-homocysteine, while 1 mM homocysteine, homoserine, or adenosine did not inhibit. The enzyme exhibited a pH optimum of 7.8 and a temperature optimum of 37°C. Activity of the isolated inositol methyltransferase was stable when stored at 4°C. |
| doi_str_mv | 10.1006/abbi.1995.1450 |
| format | Article |
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Imt1, encoding
myo-inositol
O-methyltransferase (EC 2.1.1.X) from the halophyte
Mesembryanthemum crystallinum was expressed in
Escherichia coli. The enzyme, IMT1, uses
S-adenosyl-L-methionine to methylate
myo-inositol to form D-ononitol. IMT1 with a monomeric mass of 41,000 was isolated by ammonium sulfate fractionation, gel filtration and ion exchange chromatography to apparent purity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified recombinant enzyme was identical to that encoded by the cDNA sequence. The apparent
K
m
for
S-adenosylmethionine was 0.18 mM with a
V
max of 1550 pkat/mg protein. The
K
m
for
myo-inositol was 1.32 mM. The reaction became substrate-inhibited by concentrations of
S-adenosylmethionine greater than 0.5 mM. Inositol methyltransferase was competitively inhibited 50% with 0.01 mM
S-adenosyl-homocysteine, while 1 mM homocysteine, homoserine, or adenosine did not inhibit. The enzyme exhibited a pH optimum of 7.8 and a temperature optimum of 37°C. Activity of the isolated inositol methyltransferase was stable when stored at 4°C.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1006/abbi.1995.1450</identifier><identifier>PMID: 7574673</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>ACTIVIDAD ENZIMATICA ; ACTIVITE ENZYMATIQUE ; ADN ; Amino Acid Sequence ; CLONACION ; CLONAGE ; COMPOSICION QUIMICA ; COMPOSITION CHIMIQUE ; DNA, Plant - genetics ; ESCHERICHIA COLI ; Escherichia coli - genetics ; EXPRESION GENICA ; EXPRESSION DES GENES ; INHIBICION ; INHIBITION ; INOSITOL ; Inositol - analogs & derivatives ; Inositol - chemistry ; Kinetics ; MESEMBRYANTHEMUM ; Methyltransferases - genetics ; Methyltransferases - isolation & purification ; Methyltransferases - metabolism ; Molecular Sequence Data ; Molecular Weight ; PESO MOLECULAR ; Plants - enzymology ; Plants - genetics ; POIDS MOLECULAIRE ; PURIFICACION ; PURIFICATION ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; S-Adenosylhomocysteine - metabolism ; S-Adenosylmethionine - metabolism ; Stereoisomerism ; Substrate Specificity ; TRANSFERASAS ; TRANSFERASE</subject><ispartof>Archives of biochemistry and biophysics, 1995-09, Vol.322 (1), p.183-188</ispartof><rights>1995 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-47593372d1cfba629363a8f32cb7e869bdb8ae11b14155f9b00d896ceab50d013</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0003986185714506$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7574673$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rammesmayer, G.</creatorcontrib><creatorcontrib>Pichorner, H.</creatorcontrib><creatorcontrib>Adams, P.</creatorcontrib><creatorcontrib>Jensen, R.G.</creatorcontrib><creatorcontrib>Bohnert, H.J.</creatorcontrib><title>Characterization of IMT1, myo-Inositol O-methyltransferase, from Mesembryanthemum crystallinum</title><title>Archives of biochemistry and biophysics</title><addtitle>Arch Biochem Biophys</addtitle><description>A full-length transcript,
Imt1, encoding
myo-inositol
O-methyltransferase (EC 2.1.1.X) from the halophyte
Mesembryanthemum crystallinum was expressed in
Escherichia coli. The enzyme, IMT1, uses
S-adenosyl-L-methionine to methylate
myo-inositol to form D-ononitol. IMT1 with a monomeric mass of 41,000 was isolated by ammonium sulfate fractionation, gel filtration and ion exchange chromatography to apparent purity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified recombinant enzyme was identical to that encoded by the cDNA sequence. The apparent
K
m
for
S-adenosylmethionine was 0.18 mM with a
V
max of 1550 pkat/mg protein. The
K
m
for
myo-inositol was 1.32 mM. The reaction became substrate-inhibited by concentrations of
S-adenosylmethionine greater than 0.5 mM. Inositol methyltransferase was competitively inhibited 50% with 0.01 mM
S-adenosyl-homocysteine, while 1 mM homocysteine, homoserine, or adenosine did not inhibit. The enzyme exhibited a pH optimum of 7.8 and a temperature optimum of 37°C. Activity of the isolated inositol methyltransferase was stable when stored at 4°C.</description><subject>ACTIVIDAD ENZIMATICA</subject><subject>ACTIVITE ENZYMATIQUE</subject><subject>ADN</subject><subject>Amino Acid Sequence</subject><subject>CLONACION</subject><subject>CLONAGE</subject><subject>COMPOSICION QUIMICA</subject><subject>COMPOSITION CHIMIQUE</subject><subject>DNA, Plant - genetics</subject><subject>ESCHERICHIA COLI</subject><subject>Escherichia coli - genetics</subject><subject>EXPRESION GENICA</subject><subject>EXPRESSION DES GENES</subject><subject>INHIBICION</subject><subject>INHIBITION</subject><subject>INOSITOL</subject><subject>Inositol - analogs & derivatives</subject><subject>Inositol - chemistry</subject><subject>Kinetics</subject><subject>MESEMBRYANTHEMUM</subject><subject>Methyltransferases - genetics</subject><subject>Methyltransferases - isolation & purification</subject><subject>Methyltransferases - metabolism</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>PESO MOLECULAR</subject><subject>Plants - enzymology</subject><subject>Plants - genetics</subject><subject>POIDS MOLECULAIRE</subject><subject>PURIFICACION</subject><subject>PURIFICATION</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>S-Adenosylhomocysteine - metabolism</subject><subject>S-Adenosylmethionine - metabolism</subject><subject>Stereoisomerism</subject><subject>Substrate Specificity</subject><subject>TRANSFERASAS</subject><subject>TRANSFERASE</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kL9v1DAUxy0EKkdhZUBCysTUHH5x7MQjOhV6UqsOtCuW7TxzRnFcbAcp_PVcdKduTG_4_tL7EPIe6BYoFZ-1MX4LUvIttJy-IBugUtSU9e1LsqGUslr2Al6TNzn_ohSgFc0Fueh414qObciP3UEnbQsm_1cXH6cqump_9wBXVVhivZ9i9iWO1X0dsByWsSQ9ZYdJZ7yqXIqhusOMwaRFT-WAYQ6VTUsuehz9NIe35JXTY8Z353tJHr9eP-xu6tv7b_vdl9vaMiZL3XZcMtY1A1hntGgkE0z3jjXWdNgLaQbTawQw0ALnThpKh14Ki9pwOlBgl-TTqfcpxd8z5qKCzxbHUU8Y56y6jjei6cXRuD0ZbYo5J3TqKfmg06KAqhWoWoGqFahagR4DH8_Nswk4PNvPBI_6h5PudFT6Z_JZPX6XXDQAa7g_iXj8_Y_HpLL1OFkcfEJb1BD9_3b_Af4gjdY</recordid><startdate>19950910</startdate><enddate>19950910</enddate><creator>Rammesmayer, G.</creator><creator>Pichorner, H.</creator><creator>Adams, P.</creator><creator>Jensen, R.G.</creator><creator>Bohnert, H.J.</creator><general>Elsevier Inc</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19950910</creationdate><title>Characterization of IMT1, myo-Inositol O-methyltransferase, from Mesembryanthemum crystallinum</title><author>Rammesmayer, G. ; Pichorner, H. ; Adams, P. ; Jensen, R.G. ; Bohnert, H.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-47593372d1cfba629363a8f32cb7e869bdb8ae11b14155f9b00d896ceab50d013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>ADN</topic><topic>Amino Acid Sequence</topic><topic>CLONACION</topic><topic>CLONAGE</topic><topic>COMPOSICION QUIMICA</topic><topic>COMPOSITION CHIMIQUE</topic><topic>DNA, Plant - genetics</topic><topic>ESCHERICHIA COLI</topic><topic>Escherichia coli - genetics</topic><topic>EXPRESION GENICA</topic><topic>EXPRESSION DES GENES</topic><topic>INHIBICION</topic><topic>INHIBITION</topic><topic>INOSITOL</topic><topic>Inositol - analogs & derivatives</topic><topic>Inositol - chemistry</topic><topic>Kinetics</topic><topic>MESEMBRYANTHEMUM</topic><topic>Methyltransferases - genetics</topic><topic>Methyltransferases - isolation & purification</topic><topic>Methyltransferases - metabolism</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>PESO MOLECULAR</topic><topic>Plants - enzymology</topic><topic>Plants - genetics</topic><topic>POIDS MOLECULAIRE</topic><topic>PURIFICACION</topic><topic>PURIFICATION</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>S-Adenosylhomocysteine - metabolism</topic><topic>S-Adenosylmethionine - metabolism</topic><topic>Stereoisomerism</topic><topic>Substrate Specificity</topic><topic>TRANSFERASAS</topic><topic>TRANSFERASE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rammesmayer, G.</creatorcontrib><creatorcontrib>Pichorner, H.</creatorcontrib><creatorcontrib>Adams, P.</creatorcontrib><creatorcontrib>Jensen, R.G.</creatorcontrib><creatorcontrib>Bohnert, H.J.</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Archives of biochemistry and biophysics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rammesmayer, G.</au><au>Pichorner, H.</au><au>Adams, P.</au><au>Jensen, R.G.</au><au>Bohnert, H.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of IMT1, myo-Inositol O-methyltransferase, from Mesembryanthemum crystallinum</atitle><jtitle>Archives of biochemistry and biophysics</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>1995-09-10</date><risdate>1995</risdate><volume>322</volume><issue>1</issue><spage>183</spage><epage>188</epage><pages>183-188</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><abstract>A full-length transcript,
Imt1, encoding
myo-inositol
O-methyltransferase (EC 2.1.1.X) from the halophyte
Mesembryanthemum crystallinum was expressed in
Escherichia coli. The enzyme, IMT1, uses
S-adenosyl-L-methionine to methylate
myo-inositol to form D-ononitol. IMT1 with a monomeric mass of 41,000 was isolated by ammonium sulfate fractionation, gel filtration and ion exchange chromatography to apparent purity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified recombinant enzyme was identical to that encoded by the cDNA sequence. The apparent
K
m
for
S-adenosylmethionine was 0.18 mM with a
V
max of 1550 pkat/mg protein. The
K
m
for
myo-inositol was 1.32 mM. The reaction became substrate-inhibited by concentrations of
S-adenosylmethionine greater than 0.5 mM. Inositol methyltransferase was competitively inhibited 50% with 0.01 mM
S-adenosyl-homocysteine, while 1 mM homocysteine, homoserine, or adenosine did not inhibit. The enzyme exhibited a pH optimum of 7.8 and a temperature optimum of 37°C. Activity of the isolated inositol methyltransferase was stable when stored at 4°C.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>7574673</pmid><doi>10.1006/abbi.1995.1450</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-9861 |
| ispartof | Archives of biochemistry and biophysics, 1995-09, Vol.322 (1), p.183-188 |
| issn | 0003-9861 1096-0384 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_77526286 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | ACTIVIDAD ENZIMATICA ACTIVITE ENZYMATIQUE ADN Amino Acid Sequence CLONACION CLONAGE COMPOSICION QUIMICA COMPOSITION CHIMIQUE DNA, Plant - genetics ESCHERICHIA COLI Escherichia coli - genetics EXPRESION GENICA EXPRESSION DES GENES INHIBICION INHIBITION INOSITOL Inositol - analogs & derivatives Inositol - chemistry Kinetics MESEMBRYANTHEMUM Methyltransferases - genetics Methyltransferases - isolation & purification Methyltransferases - metabolism Molecular Sequence Data Molecular Weight PESO MOLECULAR Plants - enzymology Plants - genetics POIDS MOLECULAIRE PURIFICACION PURIFICATION Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism S-Adenosylhomocysteine - metabolism S-Adenosylmethionine - metabolism Stereoisomerism Substrate Specificity TRANSFERASAS TRANSFERASE |
| title | Characterization of IMT1, myo-Inositol O-methyltransferase, from Mesembryanthemum crystallinum |
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