ABSTRACT
Kaposi sarcoma-associated herpesvirus (KSHV) is a member of the gammaherpesvirus family. It is the etiological agent of three different human cancers, Kaposi sarcoma (KS), primary effusion lymphoma (PEL) and multicentric Castleman disease. The far left end of the KSHV genome encodes a unique transmembrane glycoprotein called K1. K1 possesses the ability to transform rodent fibroblasts and block apoptosis. K1 has also been shown to activate the PI3K/Akt/mTOR pathway in different cells. Using tandem affinity purification, we identified heat shock protein 90beta (Hsp90beta) and endoplasmic reticulum-associated Hsp40 (Erdj3/DnaJB11), as cellular binding partners of K1. Interactions of K1 with Hsp90beta and Hsp40 were confirmed by co-immunoprecipitation in both directions. Furthermore, K1 also interacted with the Hsp90alpha isoform. We report that small-interfering RNAs directed against Hsp90 and Hsp40/Erdj3, as well as pharmacological inhibitors of Hsp90, dramatically reduced K1 expression, suggesting that K1 is a client protein of these chaperones. In addition, both Hsp90 and Hsp40/Erdj3 were essential for K1's anti-apoptotic function. Finally, we report that the Hsp90 inhibitors, 17-AAG and 17-DMAG, can suppress the proliferation of KSHV-positive PEL cell lines and exhibited IC(50) values of 50 nM and below.
Subject(s)
Apoptosis , Gene Expression Regulation , HSP40 Heat-Shock Proteins/metabolism , HSP90 Heat-Shock Proteins/metabolism , Herpesvirus 8, Human , Viral Proteins/metabolism , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Endoplasmic Reticulum/metabolism , Gene Knockdown Techniques , HSP40 Heat-Shock Proteins/deficiency , HSP40 Heat-Shock Proteins/genetics , HSP90 Heat-Shock Proteins/antagonists & inhibitors , HSP90 Heat-Shock Proteins/deficiency , HSP90 Heat-Shock Proteins/genetics , Humans , Lymphoma, Primary Effusion/pathology , Protein Structure, Tertiary , RNA, Small Interfering/genetics , Viral Proteins/chemistry , Viral Proteins/geneticsABSTRACT
The contents of fat and fatty acids in Callista erycina (Linnaeus), Paphia (Paratapes) undulata (Born), Meretrix meretrix (Linnaeus), Chlamys farreri (Jones et Preston) and Patinopecten yessoensis (Jay) were studied. Fat was extracted with Bligh & Dyer (B&D) method. The lipid classes were transesterified with potassium hydroxide in methanol. Fatty acid methyl esters (FAMEs) were assayed with GC-MS and polar capillary column (HP-INNOWax 30 m x 0.25 mm i.d. x 0.25 micron). GC injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. FAMEs were identified by MS library, and part by their standards. Total identified fatty acids were over 99% for all samples. Fat contents of them were all over 1% by wet samples. And ratios between omega-3PUFA and omega-6PUFA were above 2 by and large. Patinopecten yessoensis (Jay) contains more fat and the valuable fatty acids, EPA and DHA. It is suitable to use it as the source of EPA and DHA.
Subject(s)
Dietary Fats/analysis , Eicosapentaenoic Acid/analogs & derivatives , Fatty Acids/analysis , Shellfish/analysis , Animals , Eicosapentaenoic Acid/analysis , Gas Chromatography-Mass Spectrometry , Pyrones/analysisABSTRACT
The contents of twenty-three kinds of fatty acids in the lyophilized oyster that stored for 0, 15, 30, 45, 60, 75, 90 days were studied. Oyster fat was extracted from the powder by means of SFE. The extraction was performed for 40 min at a pressure of 37 MPa and a temperature of 50 degrees C with supercritical carbon dioxide containing 8% (V/V) ethanol at a flow-rate of 2 mL/min as liquid carbon dioxide, and the recovery of oyster fat by extraction was over 99%. After being extracted, fat was esterified with potassium hydroxide and methanol. Methyl esters of fatty acid were separated and determined by 30 m x 0.25 mm i.d. x 0.25 micron HP-INNOWax capillary column and MS detector. The injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. Twenty-three peaks were identified with gas chromatography/mass spectrometry, and quantified with area normalization method. The variations of contents of them were shown. During storage, the contents of saturated fatty acids and mono-unsaturated fatty acids were getting higher, and those of polyunsaturated fatty acids were getting lower. The decrease of them was gradual, and there was no special period of stability. And the stability of fatty acids in oyster related to the degree of unsaturation of them. The higher the unsaturation the lower the stable it was. After being stored for 90 days, the content of EPA decreased from 16.94% to 5.43% and that of DHA from 9.25% to 2.86%.
