{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T22:21:55Z","timestamp":1791066115047,"version":"4.1.0"},"reference-count":15,"publisher":"National Academy of Sciences","issue":"9","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2001,4,24]]},"abstract":"\n Contact sites in interaction between light-activated rhodopsin and\n transducin (T) have been investigated by using a chemically\n preactivated crosslinking reagent,\n N-<\/jats:italic>\n succinimidyl\n 3-(2-pyridyldithio)propionate. The 3\n propionyl-\n N<\/jats:italic>\n -succinimidyl group in the reagent was\n attached by a disulfide exchange reaction to rhodopsin mutants\n containing single reactive cysteine groups in the cytoplasmic loops.\n Complex formation between the derivatized rhodopsin mutants and T was\n carried out by illumination at \u03bb\u2009>\u2009495 nm. Subsequent increase in\n pH (from 6 to 7.5 or higher) of the complex resulted in crosslinking of\n rhodopsin to the T\n \u03b1<\/jats:sub>\n subunit. Crosslinking to\n T\n \u03b1<\/jats:sub>\n was demonstrated for the rhodopsin mutants K141C,\n S240C, and K248C, and the crosslinked sites in T\n \u03b1<\/jats:sub>\n were\n identified for the rhodopsin mutant S240C. The peptides carrying the\n crosslinking moiety were isolated from the trypsin-digested peptide\n mixture, and their identification was carried out by matrix-assisted\n laser desorption ionization\u2013time of flight mass spectrometry. The main\n site of crosslinking is within the peptide sequence, Leu-19\u2013Arg-28 at\n the N-terminal region of T\n \u03b1<\/jats:sub>\n .<\/jats:sub>\n The total\n results show that both the N and the C termini of T\n \u03b1<\/jats:sub>\n are\n in close vicinity to the third cytoplasmic loop of rhodopsin in the\n complex between rhodopsin and T.\n <\/jats:p>","DOI":"10.1073\/pnas.051632998","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:44:19Z","timestamp":1027680259000},"page":"4883-4887","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":85,"title":["Mapping of contact sites in complex formation between light-activated rhodopsin and transducin by covalent crosslinking: Use of a chemically preactivated reagent"],"prefix":"10.1073","volume":"98","author":[{"given":"Yoshiki","family":"Itoh","sequence":"first","affiliation":[{"name":"Departments of Biology and Chemistry, Massachusetts Institute of\r Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kewen","family":"Cai","sequence":"additional","affiliation":[{"name":"Departments of Biology and Chemistry, Massachusetts Institute of\r Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H. Gobind","family":"Khorana","sequence":"additional","affiliation":[{"name":"Departments of Biology and Chemistry, Massachusetts Institute of\r Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2001,4,24]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)48444-X"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.051632898"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6989(84)90312-2"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00512a007"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.18.6878"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2218504"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)42106-0"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi960848t"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00027a032"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1995.tb00125.x"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.52.31052"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.275.5298.381"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.3136547"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/379311a0"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1080\/07391102.2000.10506598"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.051632998","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T15:35:49Z","timestamp":1649777749000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.051632998"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,4,24]]},"references-count":15,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2001,4,24]]}},"alternative-id":["10.1073\/pnas.051632998"],"URL":"https:\/\/doi.org\/10.1073\/pnas.051632998","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,4,24]]},"assertion":[{"value":"2000-12-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2001-04-24","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}