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The resulting membrane vesicles were permeable to GDP and contained up to 3 nmol UCP\/mg protein with unchanged nucleotide binding, as compared to the mitochondria (GDP\/UCP ratio = 1.0; p<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> GDP = 6.0 at pH 7.0). UCP bound nucleotides to one type of specific binding sites, located exclusively on the cytosolic side of the mitochondrial membrane. The binding affinity of guanine nucleotides was 3\u201318\u2010times higher than that of the corresponding adenine nucleotides, when measured in membrane particles from cold\u2010acclimated rats, hamsters, and guinea pigs. The pH\u2010dependent binding affinities of GDP and ADP attained a maximum at pH 5.0\u20136.0 (p<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> GDP = 6.8, p<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> ADP = 5.8) and were decreased by a factor of 10<jats:sup>2<\/jats:sup> at pH 4.0 and pH 8.0, respectively, whereas the binding affinity of ATP was maximal at pH 4.0 (p<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub>= 7.0) and was decreased by a factor of 10<jats:sup>3<\/jats:sup> at pH 7.5. Participation of the protein binding center in nucleotide interaction with UCP in the membrane was highly pH\u2010dependent. Mg<jats:sup>2+<\/jats:sup> modified the number of binding sites engaged at a given nucleotide concentration by complex binding of nucleotides; the <jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> for Mg \u00b0 GTP<jats:sup>2\u2013<\/jats:sup> and Mg \u00b0 GDP<jats:sup>\u2212<\/jats:sup> was 20\u201350\u2010times lower than that of the free nucleotides.<\/jats:p>","DOI":"10.1111\/j.1432-1033.1994.tb19075.x","type":"journal-article","created":{"date-parts":[[2005,3,4]],"date-time":"2005-03-04T10:51:02Z","timestamp":1109933462000},"page":"971-980","source":"Crossref","is-referenced-by-count":19,"title":["Effect of pH and MgCl<sub>2<\/sub> on the binding of purine nucleotides to the uncoupling protein in membrane particles from brown fat mitochondria"],"prefix":"10.1111","volume":"223","author":[{"given":"Johannes","family":"RAFAEL","sequence":"first","affiliation":[]},{"given":"Ingelore","family":"PAMPEL","sequence":"additional","affiliation":[]},{"given":"Xuemin","family":"WANG","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2005,3,3]]},"reference":[{"key":"e_1_2_2_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1985.tb03941.x"},{"key":"e_1_2_2_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01172869"},{"key":"e_1_2_2_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(77)80748-5"},{"key":"e_1_2_2_5_1","first-page":"1756","article-title":"Microdetermination of phosphorus Anal","volume":"28","author":"Chen D. 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