engleski

Microscale thermophoresis measurements of the interaction between human dipeptidyl peptidase 3 and Kelch domain of Keap1 protein

Human dipeptidyl peptidase 3 (hDPP3) is a metallopeptidase from the M49 family of peptidases that cleaves dipeptides from the amino-termini of 3-10 long peptides in vitro. Apart from its proposed role in the final stages of proteolysis in the cell, and potential role in the regulation of blood-pressure, pain and infection through cleavage of bioactive peptides, it has been confirmed that hDPP3 is involved in the regulation of Nrf2-Keap1 signalling pathway through its interaction with Keap1 protein. Nrf2-Keap1 pathway is the main regulator of the oxidative stress response. It has been shown that hDPP3 binds Keap1, which consequently releases Nrf2 transcription factor from the complex with Keap1, and enables its translocation to the nucleus and the activation of genes coding for a series of proteins that protect the cell from the damage caused by xenobiotics and oxidative stress. Keap1 is interacting with hDPP3 through its Kelch domain. The interaction between hDPP3 and Keap1, and hDPP3 and Kelch domain of Keap1 protein has been confirmed with several methods, however the thermodynamic parameters of the interaction have not been measured. We used microscale thermophoresis (MST) to measure the dissociation constant of the hDPP3-Kelch domain complex for the wild type hDPP3, hDPP3 variant E690K, and two hDPP3 mutants found in cancer, R703C and R703H to check if the hDPP3 variant and mutants bind Kelch domain with different affinities than the wt. We performed only preliminary measurements, however, we established that it is possible to use MST to determine the thermodynamic parameters of the hDPP3-Kelch interaction.

hDPP3 ; Keap1 ; MST ; protein interaction

Объединенный научный форум, Москва, Россия, 18-22 сентября 2017 года : Международная научная конференция по биоорганической химии "XII чтения памяти академика Юрия Анатольевича Овчинникова", VIII Российский симпозиум "Белки и пептиды" : научные труды