Chromatography of isoforms of recombinant apoaequorin and method for the preparation of aequorin
Gradient elution chromatography of recombinant apoaequorin carried out in the presence of Ca 2+ revealed two isoforms of apoaequorin, reduced and oxidized, whereas in the presence of EDTA 3 isoforms were observed. In a regeneration mixture of apoaequorin, coelenterazine, EDTA, and 2-mercaptoethanol,...
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| Veröffentlicht in: | Protein expression and purification 2003-10, Vol.31 (2), p.181-187 |
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| container_title | Protein expression and purification |
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| creator | Masuda, Hiromi Takenaka, Yasuhiro Shikamoto, Yasuo Kagawa, Masayuki Mizuno, Hiroshi Tsuji, Frederick I |
| description | Gradient elution chromatography of recombinant apoaequorin carried out in the presence of Ca
2+ revealed two isoforms of apoaequorin, reduced and oxidized, whereas in the presence of EDTA 3 isoforms were observed. In a regeneration mixture of apoaequorin, coelenterazine, EDTA, and 2-mercaptoethanol, four isoforms were obtained, of which only one, aequorin, gave light with Ca
2+. A method is described for the preparation of highly pure aequorin. The aequorin was stable in solution for approximately 10 days at 4
°C and pH 7.6, and then it gradually lost activity with a half-life of about 20 days until it was almost completely inactive on day 30. |
| doi_str_mv | 10.1016/S1046-5928(03)00186-4 |
| format | Article |
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2+. A method is described for the preparation of highly pure aequorin. The aequorin was stable in solution for approximately 10 days at 4
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2+ revealed two isoforms of apoaequorin, reduced and oxidized, whereas in the presence of EDTA 3 isoforms were observed. In a regeneration mixture of apoaequorin, coelenterazine, EDTA, and 2-mercaptoethanol, four isoforms were obtained, of which only one, aequorin, gave light with Ca
2+. A method is described for the preparation of highly pure aequorin. The aequorin was stable in solution for approximately 10 days at 4
°C and pH 7.6, and then it gradually lost activity with a half-life of about 20 days until it was almost completely inactive on day 30.</description><subject>Aequorin - genetics</subject><subject>Aequorin - isolation & purification</subject><subject>Aequorin - metabolism</subject><subject>Animals</subject><subject>Apoproteins - genetics</subject><subject>Apoproteins - isolation & purification</subject><subject>Apoproteins - metabolism</subject><subject>Bioluminescence</subject><subject>Blue fluorescent protein</subject><subject>Calcium - pharmacology</subject><subject>Calcium determination</subject><subject>Chromatography</subject><subject>Coelenterazine</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Photoprotein</subject><subject>Protein Engineering</subject><subject>Protein Isoforms - genetics</subject><subject>Protein Isoforms - isolation & purification</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>Time Factors</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EgvL4BFBWCBYBvxuvEKp4SZVYAGtj7DE1auLUTpD4exJaxJLV3MW5M5qD0DHBFwQTeflEMJelULQ6w-wcY1LJkm-hCcFKlphO1faYN8ge2s_5Y4CIxGIX7REuBJZMTNDrbJFibbr4nky7-CqiL0KOPqY6jzmBjfVbaEzTFaaNBlZ9TKEpTOOKGrpFdMXAFt0CijZBa5LpQmzG5i96iHa8WWY42swD9HJ78zy7L-ePdw-z63lpmSRdya1ltLKSccOwpJRQcA6M44xOhZLcU4EV5UrQijjrGamMoopS6qn3xhN2gE7Xe9sUVz3kTtchW1guTQOxz3pKqBASqwEUa9CmmHMCr9sUapO-NMF6VKt_1OrRm8ZM_6jVfOidbA70bzW4v9bG5QBcrQEY3vwMkHS2ARoLLgwaO-1i-OfEN-lIiVM</recordid><startdate>20031001</startdate><enddate>20031001</enddate><creator>Masuda, Hiromi</creator><creator>Takenaka, Yasuhiro</creator><creator>Shikamoto, Yasuo</creator><creator>Kagawa, Masayuki</creator><creator>Mizuno, Hiroshi</creator><creator>Tsuji, Frederick I</creator><general>Elsevier Inc</general><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>20031001</creationdate><title>Chromatography of isoforms of recombinant apoaequorin and method for the preparation of aequorin</title><author>Masuda, Hiromi ; 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2+. A method is described for the preparation of highly pure aequorin. The aequorin was stable in solution for approximately 10 days at 4
°C and pH 7.6, and then it gradually lost activity with a half-life of about 20 days until it was almost completely inactive on day 30.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>14550635</pmid><doi>10.1016/S1046-5928(03)00186-4</doi><tpages>7</tpages></addata></record> |
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| ispartof | Protein expression and purification, 2003-10, Vol.31 (2), p.181-187 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Aequorin - genetics Aequorin - isolation & purification Aequorin - metabolism Animals Apoproteins - genetics Apoproteins - isolation & purification Apoproteins - metabolism Bioluminescence Blue fluorescent protein Calcium - pharmacology Calcium determination Chromatography Coelenterazine Electrophoresis, Polyacrylamide Gel Photoprotein Protein Engineering Protein Isoforms - genetics Protein Isoforms - isolation & purification Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Time Factors |
| title | Chromatography of isoforms of recombinant apoaequorin and method for the preparation of aequorin |
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