We monitored binding of ISWI to this substrate using the same native gel analysis and fluorescence anisotropy assay; the resulting data suggest that ISWI binds to this substrate with the same affinity and stoichiometry as the F10N5F substrate.
Drosophila ISWI has also been shown to bind a 35 bp DNA in a cooperative manner, indicating that more than one ISWI molecule binds to this substrate, with a reported K1/2 of 15 nM.
Our EMSA studies of the equilibrium binding of ISWI to nucleosomal substrates with very short flanking DNA, 10N5, demonstrated that ISWI binds to this substrate with a 2 1 stoichiometry.
This enzyme is not bound to the substrate, and will partition into the sugar solution after hydrolysis.
Only a certain region of the enzyme, called the active site, binds to the substrate.
Textured microspheres with small open mouths were also formed and were loosely bound to the substrate.
In this way proteins can bind to a substrate in a way that is highly specific, yet reversible.
Some DNAJs bind to substrates themselves, contributing to the substrate folding mechanism of the common HSP70 chaperones.
Natural ribozymes bind to substrate RNAs through Watson Crick base pairing, which offers the sequence-specific cleavage of substrate RNAs.
CKIε must therefore bind to substrate proteins through its kinase domain.
