{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2022,4,4]],"date-time":"2022-04-04T06:25:19Z","timestamp":1649053519946},"reference-count":0,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1989,9,15]]},"abstract":"<jats:p>A biologically active bifunctional retinoid, ethyl 14-carboxyretinoate, has been synthesized and shown to bind cellular retinoic acid (RA)-binding protein (CRABP) via its free carboxy group. We describe herein the synthesis of 14-carboxy-13-cis-retinamide-Sepharose 4B, which is an affinity matrix bearing an all-trans-RA moiety, and thus was used to purify and characterize CRABP from chick-embryo skin. An amide bond was first formed between the free carboxy group of the retinoid and a primary amino group of aminohexyl-Sepharose 4B, by reaction with carbodi-imide, and the ester group of the resin-bound retinoid was then hydrolysed in an alkaline medium. Polyacrylamide-gel electrophoresis and f.p.l.c. Superose column-chromatographic analysis demonstrated that the affinity-purified CRABP (Mr 15,000) was close to electrophoretic homogeneity (greater than 90%) and specifically interacts with RA. By using affinity gel chromatography, conversion of holo-CRABP into apo-CRABP by treatment with p-hydroxymercuribenzoate and a possible involvement of a thiol group in RA binding to CRABP were established. This affinity procedure provides several advantages: (i) 14-carboxy-13-cis-retinamide-Sepharose exhibited high efficiency and selectivity for RA-binding protein (i.e. retinol- or fatty-acid-binding proteins did not bind); (ii) the presence of the amide linkage between the ligand and the matrix makes this affinity resin relatively stable to cytosolic enzymes; and (iii) other RA-binding proteins, e.g. nuclear receptor(s), may be purified.<\/jats:p>","DOI":"10.1042\/bj2620917","type":"journal-article","created":{"date-parts":[[2015,8,10]],"date-time":"2015-08-10T21:14:08Z","timestamp":1439241248000},"page":"917-922","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Affinity purification of cellular retinoic acid-binding protein on 14-carboxy-13-<i>cis<\/i>-retinamide-sepharose 4B"],"prefix":"10.1042","volume":"262","author":[{"given":"R K","family":"Singh","sequence":"first","affiliation":[{"name":"*Kettering-Meyer Laboratories, Southern Research Institute, Birmingham, AL 35255."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B P","family":"Sani","sequence":"additional","affiliation":[{"name":"*Kettering-Meyer Laboratories, Southern Research Institute, Birmingham, AL 35255."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M I","family":"Dawson","sequence":"additional","affiliation":[{"name":"*Kettering-Meyer Laboratories, Southern Research Institute, Birmingham, AL 35255."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Y F","family":"Shealy","sequence":"additional","affiliation":[{"name":"*Kettering-Meyer Laboratories, Southern Research Institute, Birmingham, AL 35255."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/262\/3\/917\/596895\/bj2620917.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/262\/3\/917\/596895\/bj2620917.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,25]],"date-time":"2021-11-25T18:14:18Z","timestamp":1637864058000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/262\/3\/917\/24966\/Affinity-purification-of-cellular-retinoic-acid"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1989,9,15]]},"references-count":0,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1989,9,15]]}},"URL":"https:\/\/doi.org\/10.1042\/bj2620917","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[1989,9,15]]}}}