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 anonymous-2014t-1 The effects of EMIMCl on solubility, stability and aggregation of tc-rPA in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined hauke.lilie@biochemtech.uni-halle.de; tischer.alexander@mayo.edu 2014 imex curation Tischer A., Pultke H., Topf A., Auton M., Lange C., Lilie H. FEBS J. (1742-464X) Only protein-protein interactions tpa_human Tissue-type plasminogen activator PLAT protein protein human Homo sapiens Human MDAMKRGLCCVLLLCGAVFVSPSQEIHARFRRGARSYQVICRDEKTQMIYQQHQSWLRPVLRSNRVEYCWCNSGRAQCHSVPVKSCSEPRCFNGGTCQQALYFSDFVCQCPEGFAGKCCEIDTRATCYEDQGISYRGTWSTAESGAECTNWNSSALAQKPYSGRRPDAIRLGLGNHNYCRNPDRDSKPWCYVFKAGKYSSEFCSTPACSEGNSDCYFGNGSAYRGTHSLTESGASCLPWNSMILIGKVYTAQNPSAQALGLGKHNYCRNPDGDAKPWCHVLKNRRLTWEYCDVPSCSTCGLRQYSQPQFRIKGGLFADIASHPWQAAIFAKHRRSPGERFLCGGILISSCWILSAAHCFQERFPPHHLTVILGRTYRVVPGEEEQKFEVEKYIVHKEFDDDTYDNDIALLQLKSDSSRCAQESSVVRTVCLPPADLQLPDWTECELSGYGKHEALSPFYSERLKEAHVRLYPSSRCTSQHLLNRTVTDNMLCAGDTRSGGPQANLHDACQGDSGGPLVCLNDGRMTLVGIISWGLGCGQKDVPGVYTKVTNYLDWIRDNMRP plat-plat 1480554 n/a rPA 1480555 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified n/a rPA 1480555 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f5 f2b The solubility maximum of CM-rPA in 2 M EMIMCl was also measured as a function of incubation time (Fig. 2A; inset). The solubility of CM-rPA decreased over several days and after 67 days of incubation, 1.3 ± 0.1 mg mL-1 CM-rPA was detected in the soluble fraction. In addition to the solubility measurements, the state of CM-rPA in solution was analyzed by dynamic light scattering measurements (Fig. 2B). From these measurements a z-average value of 16.9 ± 0.5 nm was calculated for a sample which was incubated 19 h in 2 M EMIMCl at 4 °C. As the determined z-average values are a measure for the intensity weighed hydrodynamic diameter of particles, they indicate that CM-rPA exists as an ensemble of small aggregated protein species in solution. After an incubation of 67 days, an increase of the z-average value to 18.3 ± 0.1 was observed, indicating a shift of the aggregates to larger sizes. To test this hypothesis, we used dynamic light scattering to determine the particle size distributions (PSDs) for 54 mg mL-1 tc-rPA in 5.4 M EMIMCl as a function of time (Fig. 5). Since it was not possible to determine the exact viscosity of the EMIMCl-solution containing a certain amount of tc-rPA, the first z-average was adjusted to a diameter of 5.7 nm, which corresponded to globular, monomeric tc-rPA with a size 39.6 kDa. The measurement of the intensity weighed PSD showed a shift towards higher sizes over time. This was also reflected by the z-average diameters which increased from 5.7 nm to approximately 75 nm within 214 h. Under the assumption of a globular shape for the 75 nm particles such a z-average correlates to a hydrodynamic diameter of approximately 12 MDa indicating that tc-rPA formed soluble aggregates after unfolding in 5.4 M EMIMCl.