Binding effect and design of a competitive inhibitory peptide for HMG-CoA reductase through modeling of an active peptide backbone.

This study presents an application of two approaches in the design of constrained and unconstrained peptides in an investigation of the peptide binding effect for HMG-CoA reductase (HMGR). In previous works, hypocholesterolemic peptides isolated from soybean were determined as competitive inhibitory peptides for HMGR. Based on the modeling of an active peptide backbone in the active site of HMGR, two peptide libraries for constrained and unconstrained peptides were designed using different amino acids varying in hydrophobicity and electronic properties. Active peptides were selected by the design parameter 'V' or 'Pr', which reflects the probability of active peptide conformations for constrained and unconstrained peptides, respectively. Using peptides designed as mimics of HMGR substrates, and a combination of in vitro test and circular dichroism study, it was found that: (1) peptide binding causes an ordering of secondary structure, reflecting an increase of alpha-helical content; (2) HMGR binds the peptide without closure of the active site; and (3) peptide binding induces the protein aggregation. The GFPDGG peptide (IC(50)=1.5 microM), designed on the basis of the rigid peptide backbone, increases the inhibitory potency more than 300 times compared to the first isolated LPYP peptide (IC(50)=484 microM) from soybean. The obtained data imply the possibility of designing a highly potent inhibitory peptide for HMGR and confirm that changes of the secondary structure in the enzyme play an important role in the mechanism of HMGR inhibition.