Characteristics of the interaction mechanisms of procyanidin B1 and procyanidin B2 with protein tyrosine phosphatase-1B: Analysis by kinetics, spectroscopy methods and molecular docking

Protein tyrosine phosphatase-1B (PTP1B) is a novel and indispensable drug target for the treatment of type 2 diabetes mellitus (T2DM). Procyanidins are flavonoids that exhibit a significant hypoglycemic function. However, the potential inhibitory effects of procyanidins on PTP1B are unclear. In this study, the interaction mechanisms of PTP1B with procyanidin B1 (PB1) and procyanidin B2 (PB2) were investigated through kinetics analysis, UV-visible spectroscopy, fluorescence spectroscopy, circular dichroism spectroscopy and molecular docking. The results showed that PB1 and PB2 could inhibit the activity of PTP1B in a mixed inhibition mode, which was one of the reversible inhibition types. Multi-spectral analysis showed that PB1/PB2 formed complexes with PTP1B, which effectively quenched the intrinsic fluorescence of PTP1B based on the static mechanism. The values of the binding constants were K_S(PTP1B-PB1) = 4.06 × 10² L·mol^-1 and K_S(PTP1B-PB2) = 2.53 × 10² L·mol^-1, indicating that the binding affinity of PTP1B to PB1 was higher than that for PB2. PB1 and PB2 both changed the secondary structure of the enzyme, thereby decreasing the PTP1B activity. Thermodynamic investigations revealed that the binding of procyanidin B1 and B2 to PTP1B was spontaneous in both cases, and highlighted the key role of hydrophobic interactions. Molecular docking analysis provided further information regarding the interactions between PB1 or PB2 and the amino acid residues of PTP1B. Moreover, PB1 and PB2 were found to down-regulate the expression level of PTP1B in insulin-resistant HepG2 cells. These findings are the first to elucidate the inhibitory effects of PB1 and PB2 on PTP1B, and highlight the role of procyanidins as dietary supplements in regulating T2DM.