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During heat shock conditions, structural changes in cellular membranes may lead to cell death. Hsp90AA1 and other heat shock proteins involved in membranes are responsible for protecting membrane stabilization. However, the membrane binding mechanism of Hsp90AA1 remains largely uncharacterized. In this study, we showed Hsp90AA1 interacts with phospholipid membrane with high affinity. Using the depth-dependent fluorescence-quenching with brominated lipids, we found Hsp90AA1 penetrated 10.7 Å into the hydrocarbon core of the lipid bilayer. Circular dichroism spectra studies showed Hsp90AA1 lost part of its α-helical structures upon interaction with phospholipid membrane. By assessing binding properties of the three Hsp90AA1 domains, we found Hsp90AA1 interacted into the lipid bilayer mainly toward its C-terminus domain (CTD). Using scanning electron microscopy, we examined the protection on host cell membrane by overexpressing Hsp90AA1. The results indicated Hsp90AA1 or Hsp90AA1-CTD expressing E. coli cells exhibited better membrane integrity compared to the control after thermal treatment. The following liposome leakage assay suggested the protection of Hsp90AA1 might due to its stabilization of the membrane lipid. Collectively, the present study demonstrates Hsp90AA1 embeds into the lipid bilayer through its C-terminal domain and the Hsp90AA1-lipid association potentially has a significant function in keeping membranes stabilization during stress conditions., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Heat shock protein 90 (Hsp90) is an essential molecular chaperone with versatile functions in cell homeostatic control under both normal and stress conditions. Hsp90 has been found to be expressed on the cell surface, but the mechanism of Hsp90 association to the membrane remains obscure. In this study, the direct interaction of Hsp90 and phospholipid vesicles was characterized, and the role of Hsp90 on membrane physical state was explored. Using surface plasmon resonance (SPR), we observed a strong interaction between Hsp90 and different compositions of lipid. Hsp90 had a preference to bind with more unsaturated phospholipid species and the affinity was higher with negatively charged lipids than zwitterionic lipids. Increasing the mole fraction of cholesterol in the phospholipid led to a decrease of binding affinity to Hsp90. Circular dichroism (CD) spectroscopy of Hsp90 in PC membranes showed more α-helix structure than in aqueous buffer. The differential scanning calorimeter (DSC) and fluorescence polarization results showed Hsp90 could affect the transition temperature and fluidity of the bilayer. We postulate from these results that the association between Hsp90 and membranes may involve both electrostatic and hydrophobic force, and constitute a possible mechanism that modulates membrane lipid order during thermal fluctuations., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The antioxidative role of Hsp90 purified from duck muscle on phospholipid was investigated in this study. Lipid oxidation was assessed by ferric thiocyanate method, and particle size, zeta potential measurements and transmission electron microscopy were employed to detect physical and chemical changes during oxidation of liposomes. Increase of the peroxidation products, decrease of the particle size and disruption of the spherical unity of the liposomes were detected when liposome was exposed to hydrogen peroxide (H 2 O 2 ) oxidation, and Hsp90 was shown to protect liposome against oxidation. To further test the protection of Hsp90 on cell membrane, the Hsp90 transformed E. coli cell model was built. The Hsp90-containing E. coli showed higher survival and better membrane integrity as compared to the control upon H 2 O 2 exposure. It is expected that Hsp90 plays an important role in the prevention of phospholipids oxidation in meat and has potential use as a food antioxidant., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Phospholipid hydrolysis, as the main stage and cause of phopholipid degradation, is generally attributed to phospholipases. In this study, the changes of phospholipase A₂ (PLA₂) and C (PLC) activities, neutral lipid, free fatty acids and phospholipids in dry-cured duck leg muscles during processing, were examined. The composition of free fatty acids and neutral lipids increased significantly (P<0.05) with extension of processing time while the phospholipids composition decreased. The PLA₂ and PLC activity decreased in the final product, but retained 83.70% and 86.78% of their initial activities, respectively. The relative activities of both PLA₂ and PLC highly correlated with the decline of phospholipids and the increase of free fatty acids. High correlations were also obtained between the relative activities of PLC and the increase of neutral lipid (P<0.01). All these results suggest that PLA₂ and PLC contribute to the degradation of intramuscular phospholipids during the processing of dry-cured duck., (Copyright © 2013. Published by Elsevier Ltd.)

Endogenous phospholipase A 2 (PLA 2) plays an important role in phospholipids degradation during cured meat products manufacturing. The present study was undertaken to reveal more information about the endogenous PLA 2 in muscles and its role in degradation of intramuscular phospholipids. With the catalytic domain of pork calcium-independent PLA 2 (iPLA 2 cd), impacts of physic-chemical factors on the activity were investigated and substrate specificity of the enzyme were tested respectively. The optimum temperature and pH of pork iPLA 2 cd were 40 °C and 7.5, respectively. The iPLA 2 cd could be stimulated by adequate contents of NaCl and ATP, and inhibited by CaCl 2 and NaNO 2. For native phospholipids, the iPLA 2 cd was of a little higher affinity towards phosphatidylcholine (PC) than phosphatidylethanolamine (PE), phosphoserine (PS) and phosphatidylinositol (PI). The iPLA 2 cd could preferentially hydrolyze peroxidized PC over the native PC. The results would help better understand the degradation of phospholipids and the role played by endogenous enzymes during meat products manufacturing. [ABSTRACT FROM AUTHOR]