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 http://mint.bio.uniroma2.it/mint dub-2016-1 in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Dubé A., Leggiadro C., Ewart KV. vewart@dal.ca FEBS letters 2016 Only protein-protein interactions imex curation imex curation dub-2016-2 in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Dubé A., Leggiadro C., Ewart KV. vewart@dal.ca FEBS letters 2016 Only protein-protein interactions imex curation imex curation dub-2016-3 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Dubé A., Leggiadro C., Ewart KV. vewart@dal.ca FEBS letters 2016 Only protein-protein interactions imex curation imex curation anpa_pseam Ice-structuring protein A Antifreeze protein A HPLC6 protein protein Winter flounder Pseudopleuronectes americanus Pleuronectes americanus Winter flounder MALSLFTVGQLIFLFWTMRITEASPDPAAKAAPAAAAAPAAAAPDTASDAAAAAALTAANAKAAAELTAANAAAAAAATARG anpa_pseam-anpa_pseam-1 1939149 N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f3 f6 f7 f8 f9 Equilibrium freezing of 15 mm wflAFP-6 in the presence of an ice seed resulted in strong ThT fluorescence (Fig. 3), while spontaneously nucleated rapid phase change in the absence of an ice seed and maintenance of a low temperature without freezing each produced far weaker signals similar to those of control samples held at 4 °C. Thus, it is the specific binding of wflAFP-6 to the ice surface that favours its transition to an amyloid structure, rather than temperature or phase changes that occur during the freezing process. anpa_pseam-anpa_pseam-2 1939150 N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f4 When examined by TEM, the samples that had undergone ice interaction (ice-seeded freezing) revealed fibrils (Fig. 4), whereas none were present in the control samples that were held unfrozen at 4 °C. anpa_pseam-anpa_pseam-3 1939151 N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified N/A 1939152 unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f5 The protein size profile analysed by size exclusion chromatography also shifted in ice interaction samples compared with unfrozen (4 °C) controls (Fig. 5). Early-eluting protein with relative masses > 67 kDa (the largest marker) increased sharply (peaks 1 and 2), whereas the major peak (4) corresponding to a mass of 10 kDa, which is the predicted mass of monomeric wflAFP-6 in size exclusion chromatography due to its extended structure [21, 22], was diminished. These results suggest extensive protein association. A 32-kDa peak (3) was also present in both samples, which may represent modest oligomerization under both conditions. The size exclusion and TEM results are consistent with the ThT fluorescence indicating amyloid formation.