|Compound Screen Assay, Human CENTG1 - BioAssay Summary
The apparent binding of compounds to Human CENTG1 has been measured using differential scanning fluorimetry (DSF) technique. ..more
BioActive Compounds: 2
The apparent binding of compounds to Human CENTG1 has been measured using differential scanning fluorimetry (DSF) technique.
In this approach, the unfolding of the protein is monitored via a fluorescent hydrophobicity-sensing dye: SYPRO orange (Invitrogen). The hyperbolic curve of increase in fluorescence intensity versus temperature shows the unfolding process. The point of inflexion of this trace provides the measured parameter known as Tm. A difference in Tm between a control without the compound and an experiment with the compound present is called a TmShift. Positive shifts indicate that the protein is more stable in the presence of the compound, indicating that the compound is bound. Since it is not possible to quantitatively compare TmShift values from different experiments containing different components, we mark positive shifts of more than 3 degrees celcius as having a PubChem assay score of 100, and all other shifts with a score of 0.
The family of small GTPases play major roles in many cellular processes including cytoskeleton regulation, membrane trafficking, signal transduction and nuclear import. They perform these functions by existing in two forms an active (GTP bound) state and an inactive (GDP bound) state. The reversible cycling between these two states is important in the signalling process and is modified by a number of regulatory proteins including guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs). Centaurin gamma 1 (CENTG1) is a multi-domain protein that based on sequence alignments contains both a GTPase domain and a GAP domain. Here we have determined the structure of the GTPase domain of CENTG1. The overall fold resembles that of known GTPase structures. However, in striking contrast to all other structures of small GTPases no nucleotide was present in the potential nucleotide binding site. As it remains unclear to the ability of this domain to hydrolyse GTP it has been labelled a GTPase-like domain.
Data Table (Concise)