Isolation, purification, and study of properties of recombinant hepsin from Escherichia coli.
A recombinant hepsin-producing strain of Escherichia coli was obtained and the conditions for hepsin expression in a bacterial system were optimized. To study the physicochemical properties of the enzyme, a procedure for purification of active recombinant hepsin using metal-chelate affinity chromatography and ion-exchange chromatography was developed. The interaction of recombinant hepsin with various peptide substrates is characterized. The dose-dependent inhibition of the recombinant hepsin enzyme activity by anthralin in vitro and an increase in the hepsin enzymatic activity in the presence of resveratrol were revealed.
Raevskaya
A A
AA
Lomonosov Moscow State University, Moscow, Russia. araevskaia@gmail.com
Kuznetsova
E M
EM
Savvateeva
M V
MV
Severin
S E
SE
D000875:Anthralin / Q000494:pharmacology; D045744:Cell Line, Tumor; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000077185:Resveratrol; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013267:Stilbenes / Q000494:pharmacology; D013379:Substrate Specificity
D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D013267:Stilbenes; D012697:Serine Endopeptidases; C055645:hepsin; D000077185:Resveratrol; D000875:Anthralin