Title
Characterization of molecular properties and regulatory pathways of CrustinPm1 and CrustinPm7 from the black tiger shrimp Penaeus monodon
Author
Sopacha Arayamethakorn, Premruethai Supungul, Anchalee Tassanakajon, Kuakarun Krusong
Year
2017
Journal
Developmental & Comparative Immunology
Abstract
CrustinPm1 and crustinPm7 are the two most abundant isoforms of crustins identified from the hemocytes of the black tiger shrimp, Penaeus monodon. CrustinPm1 inhibits only Gram-positive bacteria, while crustinPm7 acts against both Gram-positive and Gram-negative bacteria. This work aims to characterize the molecular properties of recombinant crustinPm1 and crustinPm7, and the regulatory pathways of these two crustins. Circular dichroism spectroscopy revealed that crustinPm1 contained 40.81% alpha-helix and 22.34% beta-sheet, whereas crustinPm7 is made up of 32.86% alpha-helix and 27.53% beta-sheet. CrustinPm1 and crustinPm7 bound to phosphatidic acid (PA) with positive cooperativity of Hill slope (H) > 2, indicating that at least two molecules of crustins bind with one PA molecule. It is worth noting that both crustins bound to PA with significantly higher affinity than to lipoteichoic acid (LTA) and lipopolysaccharide (LPS). We speculate that crustin might also achieve antimicrobial activity by targeting PA, a signaling lipid. Regulatory pathways of crustinPm1 and crustinPm7 were investigated by knockdown of PmRelish and PmMyD88. This study demonstrated that crustinPm1 is mediated through the Toll signaling pathway, while crustinPm7 is regulated via both Toll and Imd pathways.
Instrument
J-715
Keywords
Circular dichroism, Secondary structure, Biochemistry