MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Cu(I)- and proton-binding properties of the first N-terminal soluble domain of Bacillus subtilis CopA. Cu(I)- and proton-binding properties of the first N-terminal soluble domain of Bacillus subtilis CopA. FEBS J. (1742-464X) 2012 imex curation Zhou L., Singleton C., Hecht O., Moore GR., Le Brun NE. Only protein-protein interactions in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Zhou L., Singleton C., Hecht O., Moore GR., Le Brun NE. FEBS J. (1742-464X) 2012 Only protein-protein interactions imex curation Cu(I)- and proton-binding properties of the first N-terminal soluble domain of Bacillus subtilis CopA. Cu(I)- and proton-binding properties of the first N-terminal soluble domain of Bacillus subtilis CopA. FEBS J. (1742-464X) 2012 imex curation Zhou L., Singleton C., Hecht O., Moore GR., Le Brun NE. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Zhou L., Singleton C., Hecht O., Moore GR., Le Brun NE. FEBS J. (1742-464X) 2012 Only protein-protein interactions imex curation copa_bacsu Copper-exporting P-type ATPase Copper-exporting P-type ATPase A copA BSU33500 yvgX Cu(+)-exporting ATPase protein protein bacsu Bacillus subtilis MSEQKEIAMQVSGMTCAACAARIEKGLKRMPGVTDANVNLATETSNVIYDPAETGTAAIQEKIEKLGYHVVTEKAEFDIEGMTCAACANRIEKRLNKIEGVANAPVNFALETVTVEYNPKEASVSDLKEAVDKLGYKLKLKGEQDSEAAAKKKEERKQTARLIFSAVLSFPLLWAMVSHFTFTSFIWVPDIFLNPWMQFALATPVQFLIGWPFYVGAYKALRNKSANMDVLVALGTTAAYAYSLYLTFQSIGSHGHTDGLYYETSAILLTLILLGKLFETKAKGRSSDAIKKLMKLQAKTATVVRDGQEQIIPIDEVLVNDIVYVKPGERIPVDGEVVEGRSAVDESMITGESLPVDKNPGDSVTGSTVNANGFLKIKAVNVGKDTALSHIIKIVEEAQGSKAPIQRLADQISGIFVPIVLGIAVLTFLIWYLWAAPGDFAEAISKFIAVLVIACPCALGLATPTSIMAGSGRAAEFGILFKGGEHLEKTHRLDTIVLDKTGTVTNGKPRLTDAIPFGRFEEKDLLQFAAAAETGSEHPLGEAIIAGVKDKGLEIPKLTRFEAKVGAGILAEAGGKSILVGTRKLMESEQVEHGALLAQMEELEAEGKTVMLVSIDGEAAGLVAVADTIKDTSRKAVARLKELGLDVIMMTGDNRRTAEAIAKEAGIANIIAEVLPEQKAAEIARLQKEGRQTAMVGDGINDAPALATADIGMAIGTGTDIAMETADITLIRGDLNSIADAIRMSRLTMKNIKQNLFWALGYNSLGIPIAALGFLAPWIAGAAMAFSSVSVVLNALRLQKVK 8D6EC2B203E11E38 copa-copa 2 CopA 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified CopA 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f4 mint A number of lower intensity band shoulders were also observed at lower elution volumes, suggesting that a proportion of CopAa was present as higher molecular mass forms, perhaps as disulfide bonded oligomers. The chromatograms for Cu(I)-bound CopAa revealed very different elution profiles. CopAa containing 0.5 Cu(I)/protein (Figure 4) gave a main elution peak volume of 11 ml, with shoulders at 13.6 and 14.6 ml, indicating that most CopAa was present as a significantly higher molecular weight form, with smaller proportions present as monomer and perhaps dimer forms copa-1 1 CopA 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f5 mint Fits of data for apo-CopAa, see Figure 5A for data at 28,000 rpm, gave an average molecular mass of 8.2 ± 0.2 kDa, slightly higher than the calculated molecular mass of CopAa, (7.6 kDa). Thus, apo-CopAa is a monomer, as concluded from analytical gel filtration chromatography (see above). At a loading of 0.5 Cu(I)/CopAa the molecular mass increased significantly up 20.5 ± 0.5 kDa. Clearly, the protein undergoes Cu(I)- mediated association. 20 kDa is closest to a trimeric form of CopAa, but reference to the analytical gel filtration data, which showed two major species, suggests that, at 0.5 Cu(I)/CopAa, there is a mixture, which may be in slow equilibrium, between likely tetramer and monomer species, with an average molecular mass of 20 kDa. At a loading of 2 Cu(I)/CopAa, fitting of the data gave a molecular weight of 9.8 ± 0.6 kDa. Again, with reference to analytical gel filtration data, it can be concluded that, at this level of Cu(I), the protein is present principally as an equilibrium mixture of monomer and dimer, with the monomer as the major species