{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Krishnan K"],"funding":["Taylor Family Institute for Innovative Psychiatric Research, Washington University in St. Louis","NIMH NIH HHS","NHLBI NIH HHS","National Heart, Lung, and Blood Institute","National Institute of Mental Health","National Institute of General Medical Sciences","NIGMS NIH HHS"],"pagination":["1493-1507"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC8740335"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["16(8)"],"pubmed_abstract":["Aliphatic diazirine analogues of cholesterol have been used previously to elaborate the cholesterol proteome and identify cholesterol binding sites on proteins. Cholesterol analogues containing the trifluoromethylphenyl diazirine (TPD) group have not been reported. Both classes of diazirines have been prepared for neurosteroid photolabeling studies and their combined use provided information that was not obtainable with either diazirine class alone. Hence, we prepared cholesterol TPD analogues and used them along with previously reported aliphatic diazirine analogues as photoaffinity labeling reagents to obtain additional information on the cholesterol binding sites of the pentameric <i>Gloeobacter</i> ligand-gated ion channel (GLIC). We first validated the TPD analogues as cholesterol substitutes and compared their actions with those of previously reported aliphatic diazirines in cell culture assays. All the probes bound to the same cholesterol binding site on GLIC but with differences in photolabeling efficiencies and residues identified. Photolabeling of mammalian (HEK) cell membranes demonstrated differences in the pattern of proteins labeled by the two classes of probes. Collectively, these date indicate that cholesterol photoaffinity labeling reagents containing an aliphatic diazirine or TPD group provide complementary information and will both be useful tools in future studies of cholesterol biology."],"journal":["ACS chemical biology"],"pubmed_title":["Validation of Trifluoromethylphenyl Diazirine Cholesterol Analogues As Cholesterol Mimetics and Photolabeling Reagents."],"pmcid":["PMC8740335"],"funding_grant_id":["T32 HL134635","R01 GM108799","F31 HL142167","R01 HL067773","R01 MH110550"],"pubmed_authors":["Covey DF","Chen ZW","Ory DS","Krishnan K","Feltes M","Gale S","Evers AS","Sugasawa Y","Reichert DE","Schaffer JE","Qian M","Wang L"],"additional_accession":[]},"is_claimable":false,"name":"Validation of Trifluoromethylphenyl Diazirine Cholesterol Analogues As Cholesterol Mimetics and Photolabeling Reagents.","description":"Aliphatic diazirine analogues of cholesterol have been used previously to elaborate the cholesterol proteome and identify cholesterol binding sites on proteins. Cholesterol analogues containing the trifluoromethylphenyl diazirine (TPD) group have not been reported. Both classes of diazirines have been prepared for neurosteroid photolabeling studies and their combined use provided information that was not obtainable with either diazirine class alone. Hence, we prepared cholesterol TPD analogues and used them along with previously reported aliphatic diazirine analogues as photoaffinity labeling reagents to obtain additional information on the cholesterol binding sites of the pentameric <i>Gloeobacter</i> ligand-gated ion channel (GLIC). We first validated the TPD analogues as cholesterol substitutes and compared their actions with those of previously reported aliphatic diazirines in cell culture assays. All the probes bound to the same cholesterol binding site on GLIC but with differences in photolabeling efficiencies and residues identified. Photolabeling of mammalian (HEK) cell membranes demonstrated differences in the pattern of proteins labeled by the two classes of probes. Collectively, these date indicate that cholesterol photoaffinity labeling reagents containing an aliphatic diazirine or TPD group provide complementary information and will both be useful tools in future studies of cholesterol biology.","dates":{"release":"2021-01-01T00:00:00Z","publication":"2021 Aug","modification":"2022-02-11T14:59:25.118Z","creation":"2022-02-11T14:59:25.118Z"},"accession":"S-EPMC8740335","cross_references":{"pubmed":["34355883"],"doi":["10.1021/acschembio.1c00364"]}}