Purification and Inhibitory Mechanism of a Prolyl Endopeptidase Inhibitor from the Skeletal Muscle of Blue Scad (Decapterus maruadsi)

Abstract

Abstract: In the present study, the inhibitory effect and mechanism of an endogenous prolyl endopeptidase inhibitor (PEPI) on prolyl endopeptidase (PEP) were investigated using marine fish blue scad (Decapterus maruadsi) as raw material. PEPI was purified from the skeletal muscle of blue scad by thermal precipitation, ammonium sulfate fractionation and a series of column chromatographies including DEAE-Sepharose, Sephacryl S-200 HR and HiTrap Q HP. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) showed that the molecular weight of PEPI was about 10 kDa. It is thermal stable and tolerance to pH change. PEPI is a specific inhibitor to serine proteinase PEP and it works in a reversible competitive manner with inhibitory constant Ki of 0.34 μmol/L. A significant increase in α-helix and random coil and decrease in β-sheet were detected after formation of the PEP-PEPI complex. Quite possibly, conformation change of PEP is the major reason for enzymatic activity inhibition. Analysis of the inhibitory mechanism of PEPI to PEP would provide a theoretical basis for revealing the physiological function of PEP.

Key words: blue scad, prolyl endopeptidase, endogenous inhibitor, inhibitory kinetic, secondary structure

CLC Number:

TS254.4

chan zhong Liu Guang-Ming. Purification and Inhibitory Mechanism of a Prolyl Endopeptidase Inhibitor from the Skeletal Muscle of Blue Scad (Decapterus maruadsi)[J]. FOOD SCIENCE, 0, (): 0-0.

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