Rational design of an N-terminal cysteine-containing tetrapeptide that inhibits tyrosinase and evaluation of its mechanism of action

Anupong Joompang; Preeyanan Anwised; Sompong Klaynongsruang; Lapatrada Taemaitree; Anuwat Wanthong; Kiattawee Choowongkomon; Sakda Daduang; Somporn Katekaew; Nisachon Jangpromma

doi:10.1016/j.crfs.2023.100598

Rational design of an N-terminal cysteine-containing tetrapeptide that inhibits tyrosinase and evaluation of its mechanism of action

Curr Res Food Sci. 2023 Sep 21:7:100598. doi: 10.1016/j.crfs.2023.100598. eCollection 2023.

Authors

Anupong Joompang^{1

2}, Preeyanan Anwised², Sompong Klaynongsruang^{1

2}, Lapatrada Taemaitree³, Anuwat Wanthong^{2

4}, Kiattawee Choowongkomon⁵, Sakda Daduang^{2

6}, Somporn Katekaew^{1

2}, Nisachon Jangpromma^{1

2}

Affiliations

¹ Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand.
² Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand.
³ Department of Integrated Science, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand.
⁴ Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham, 44150, Thailand.
⁵ Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand.
⁶ Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.

Abstract

There has been a resurgence of interest in bioactive peptides as therapeutic agents. This is particularly interesting for tyrosinase, which can be inhibited by thiol-containing peptides. This work demonstrates that an N-terminal cysteine-containing tetrapeptide can be rationally designed to inhibit tyrosinase activity in vitro and in cells. The tetrapeptide cysteine (C), arginine (R), asparagine (N) and leucine (L) or CRNL is a potent inhibitor of tyrosinase activity with an IC₅₀ value of 39.62 ± 6.21 μM, which is comparable to currently used tyrosinase inhibitors. Through structure-activity studies and computational modeling, we demonstrate the peptide interacts with the enzyme via electrostatic (R with E322), hydrogen bonding (N with N260) and hydrophobic (L with V248) intermolecular interactions and that a combination of these is required for potent activity. Moreover, copper chelating activity might be one of the mechanisms of tyrosinase inhibition by CRNL. Kinetic studies show that tetrapeptide is a competitive inhibitor with two-step irreversible inhibition. In addition, CRNL had no toxicity and could reduce melanin levels in the murine melanoma cell line (B16F1). Overall, CRNL is a very promising candidate for hyperpigmentation treatment.

Keywords: Depigmentation; Enzyme kinetics; Modified peptide; Molecular docking; Tetrapeptide; Tyrosinase inhibitor.