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We also identified specific non-catalytic residues that are involved in stabilizing the substrate at the optimal pH for enzyme activity.
The phosphate moiety rapidly associates with R45 in the apex whose guanidinium group acts like a "fork" stabilizing the substrate in its binding site.
Additional changes in the active site enhance the rate of phosphotransfer by stabilizing the substrate in the transition state, thus enhancing the reactivity of Cdc6 (73 75).
The unprecedented substrate binding mode and cleavage pathway for phenylpyruvate-type structures at the 3-His center of Dke1 suggest that the metal binding motif may contribute to stabilizing the substrate ligand in the correct ionization state.
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Deubiquitinating enzymes (DUBs) serve to remove Ub from ubiquitinated proteins, thus stabilizing the substrates.
Reciprocally, substrate binding induces ATP hydrolysis, "closing" the SDB and thus stabilizing the substrate-HSP70 complex [ 27, 28].
The threonine in position 254 is also found as either threonine (usually) or serine (more rarely) in homologs; this site is equivalent to T160 in human Cdk2, where phosphorylation of this residue dramatically increases CDK catalytic activity, apparently stabilizing the substrate-binding site by forming a network of hydrogen bonds with surrounding side chains [ 71].
Furthermore, His194 cannot form part of a His-Ser catalytic dyad and only stabilizes the substrate.
The hydrophobic wall comprised of I167 and F212 is adjacent to the hydrophobic side of the substrate, which stabilizes the substrate binding.
Therefore, the mutation I211A adjusted the local structure near the binding pocket and stabilizes the substrate binding with the specificity subsite of OPHs.
Inhibitor binding not only stabilized the substrate binding loop but also revealed that the substrate binding tunnel has an overall shape of "U".
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