{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T10:25:33Z","timestamp":1761560733853},"reference-count":29,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2001,12,25]],"date-time":"2001-12-25T00:00:00Z","timestamp":1009238400000},"content-version":"vor","delay-in-days":816,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[1999,10]]},"abstract":"<jats:p>The early endosomal autoantigen EEA1 is essential for early endosomal membrane fusion. It binds to endosomes via a C\u2010terminal domain (EEA1\u2010CT). To identify proteins interacting with EEA1\u2010CT, we screened a human brain library in the yeast two\u2010hybrid system. Fourteen clones reacted strongly with EEA1\u2010CT. Sequencing of these clones revealed that they all contained the ORF of the small GTPase, Rab5b. Further two\u2010hybrid analysis suggested that Rab5b also interacts with the N\u2010terminus of EEA1 (EEA1\u2010NT). The interaction of both EEA1\u2010CT and EEA1\u2010NT with Rab5b was confirmed biochemically, and was found to be GTP dependent. Confocal immunofluorescence microscopy indicated that EEA1 colocalizes with Rab5b on early endosomes. Although EEA1\u2010CT and EEA1\u2010NT interacted strongly with wild\u2010type Rab5b in the two\u2010hybrid system, we detected no interaction with wild\u2010type Rab5a, even though GTPase\u2010deficient mutants of both Rab5a and Rab5b interacted equally well with EEA1. This difference could not be explained by differences in intrinsic GTPase activities, as these were found to be very similar. Instead, we speculate that yeast may contain a GTPase\u2010activating protein (GAP) activity that stimulates Rab5a but not Rab5b. In contrast, pig brain cytosol was found to contain a GAP activity that stimulates the GTPase activity of Rab5b in preference to that of Rab5a. These data provide evidence that EEA1 interacts with both Rab5a and Rab5b, and that the GTPase activities of the two proteins are differentially regulated <jats:italic>in vivo<\/jats:italic>.<\/jats:p>","DOI":"10.1046\/j.1432-1327.1999.00743.x","type":"journal-article","created":{"date-parts":[[2003,3,11]],"date-time":"2003-03-11T17:35:16Z","timestamp":1047404116000},"page":"361-366","source":"Crossref","is-referenced-by-count":52,"title":["Direct interaction of EEA1 with Rab5b"],"prefix":"10.1111","volume":"265","author":[{"given":"Judy","family":"Callaghan","sequence":"first","affiliation":[]},{"given":"Susan","family":"Nixon","sequence":"additional","affiliation":[]},{"given":"Cecilia","family":"Bucci","sequence":"additional","affiliation":[]},{"given":"Ban\u2010Hock","family":"Toh","sequence":"additional","affiliation":[]},{"given":"Harald","family":"Stenmark","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2001,12,25]]},"reference":[{"key":"e_1_2_7_2_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj3360271"},{"key":"e_1_2_7_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2499930"},{"key":"e_1_2_7_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(91)90316-Q"},{"key":"e_1_2_7_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80380-3"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/17618"},{"key":"e_1_2_7_7_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.22.13503"},{"key":"e_1_2_7_8_2","doi-asserted-by":"publisher","DOI":"10.1006\/clin.1997.4455"},{"key":"e_1_2_7_9_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.39.24048"},{"key":"e_1_2_7_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/28879"},{"key":"e_1_2_7_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/28767"},{"key":"e_1_2_7_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.21.9578"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90120-5"},{"key":"e_1_2_7_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(95)00477-Q"},{"key":"e_1_2_7_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90307-C"},{"key":"e_1_2_7_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00340712"},{"key":"e_1_2_7_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-9525(96)80008-4"},{"key":"e_1_2_7_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(83)01013-7"},{"key":"e_1_2_7_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.5.12.3610"},{"key":"e_1_2_7_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(95)00843-X"},{"key":"e_1_2_7_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(95)57021-7"},{"key":"e_1_2_7_22_2","doi-asserted-by":"crossref","unstructured":"Simonsen A. 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