Affinity alkylation of bacterial delta 5-3-ketosteroid isomerase. Identification of the amino acid modified by steroidal 17 beta-oxiranes
The two steroidal 17 beta-oxiranes, spiro-17 beta-oxiranyl-delta 4-androsten-3-one (4 beta) and spiro-17 beta-oxiranylestra-1,3,5(10),6,8-pentaene-3-ol (5 beta) are active site-directed irreversible inhibitors of bacterial delta 5-3-ketosteroid isomerase. For each inhibitor, a stoichiometry of one m...
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| Veröffentlicht in: | The Journal of biological chemistry 1983-01, Vol.258 (2), p.909-915 |
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| container_issue | 2 |
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| container_title | The Journal of biological chemistry |
| container_volume | 258 |
| creator | Kayser, R H Bounds, P L Bevins, C L Pollack, R M |
| description | The two steroidal 17 beta-oxiranes, spiro-17 beta-oxiranyl-delta 4-androsten-3-one (4 beta) and spiro-17 beta-oxiranylestra-1,3,5(10),6,8-pentaene-3-ol (5 beta) are active site-directed irreversible inhibitors of bacterial delta 5-3-ketosteroid isomerase. For each inhibitor, a stoichiometry of one molecule of steroid to one enzyme subunit was found. The inhibited enzyme was denatured and subjected to digestion by trypsin. The tryptic maps show two distinct steroid-bound peptides for both 4 beta- and 5 beta-inhibited isomerase. In each case, the two modified peptides are derived from residues 14 to 45 of the isomerase. Each of the steroid-bound peptides of the 4 beta-inhibited enzyme was subjected to further proteolytic digestion and the site of steroid attachment was found to be Asp-38 in each of the inactivation products. These results are interpreted to indicate that "backwards binding" is an important feature of the binding of steroids to delta 5-3-ketosteroid isomerase. |
| doi_str_mv | 10.1016/S0021-9258(18)33137-5 |
| format | Article |
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Identification of the amino acid modified by steroidal 17 beta-oxiranes</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Kayser, R H ; Bounds, P L ; Bevins, C L ; Pollack, R M</creator><creatorcontrib>Kayser, R H ; Bounds, P L ; Bevins, C L ; Pollack, R M</creatorcontrib><description>The two steroidal 17 beta-oxiranes, spiro-17 beta-oxiranyl-delta 4-androsten-3-one (4 beta) and spiro-17 beta-oxiranylestra-1,3,5(10),6,8-pentaene-3-ol (5 beta) are active site-directed irreversible inhibitors of bacterial delta 5-3-ketosteroid isomerase. For each inhibitor, a stoichiometry of one molecule of steroid to one enzyme subunit was found. The inhibited enzyme was denatured and subjected to digestion by trypsin. The tryptic maps show two distinct steroid-bound peptides for both 4 beta- and 5 beta-inhibited isomerase. In each case, the two modified peptides are derived from residues 14 to 45 of the isomerase. Each of the steroid-bound peptides of the 4 beta-inhibited enzyme was subjected to further proteolytic digestion and the site of steroid attachment was found to be Asp-38 in each of the inactivation products. These results are interpreted to indicate that "backwards binding" is an important feature of the binding of steroids to delta 5-3-ketosteroid isomerase.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(18)33137-5</identifier><identifier>PMID: 6822514</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acids - analysis ; Androstenes - pharmacology ; Binding Sites ; Chromatography, High Pressure Liquid ; Estrenes - pharmacology ; Isomerases - antagonists & inhibitors ; Peptide Fragments - analysis ; Steroid Isomerases - antagonists & inhibitors ; Time Factors ; Trypsin - metabolism</subject><ispartof>The Journal of biological chemistry, 1983-01, Vol.258 (2), p.909-915</ispartof><rights>1983 © 1983 ASBMB. 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Identification of the amino acid modified by steroidal 17 beta-oxiranes</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The two steroidal 17 beta-oxiranes, spiro-17 beta-oxiranyl-delta 4-androsten-3-one (4 beta) and spiro-17 beta-oxiranylestra-1,3,5(10),6,8-pentaene-3-ol (5 beta) are active site-directed irreversible inhibitors of bacterial delta 5-3-ketosteroid isomerase. For each inhibitor, a stoichiometry of one molecule of steroid to one enzyme subunit was found. The inhibited enzyme was denatured and subjected to digestion by trypsin. The tryptic maps show two distinct steroid-bound peptides for both 4 beta- and 5 beta-inhibited isomerase. In each case, the two modified peptides are derived from residues 14 to 45 of the isomerase. Each of the steroid-bound peptides of the 4 beta-inhibited enzyme was subjected to further proteolytic digestion and the site of steroid attachment was found to be Asp-38 in each of the inactivation products. These results are interpreted to indicate that "backwards binding" is an important feature of the binding of steroids to delta 5-3-ketosteroid isomerase.</description><subject>Amino Acids - analysis</subject><subject>Androstenes - pharmacology</subject><subject>Binding Sites</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Estrenes - pharmacology</subject><subject>Isomerases - antagonists & inhibitors</subject><subject>Peptide Fragments - analysis</subject><subject>Steroid Isomerases - antagonists & inhibitors</subject><subject>Time Factors</subject><subject>Trypsin - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1983</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1uEzEUhS0EKmnhESqZDaILF_-MM_YKVRUtlSqxKEjsLP_cIaYz42I7hTxC3xqnibLFG0u-37k-OgehU0bPGWXLj3eUckY0l-oDU2dCMNET-QItGFWCCMl-vESLA_IaHZfyi7bTaXaEjpaKc8m6BXq6GIY4x7rBdrzfjLbGNOM0YGd9hRztiAOM1WJJBLmHmkp7TTHgWNIE2RY4xzcB5hqH6A_iugJspzgnbH1jpxTaGAJ2G7zXt72sxw6qJelvzHaG8ga9GuxY4O3-PkHfrz5_u_xCbr9e31xe3BLfCVGJHLq-U5xaL7heSuaFFszyQXTAGWcUehWc0z10vdCDE6HRundaBwGUei1O0Pvd3oecfq-hVDPF4mEcm4m0LkZRsVRa0wbKHehzKiXDYB5ynGzeGEbNtgLzXIHZ5muYMs8VGNl0p_sP1m6CcFDtM2_zd7v5Kv5c_YkZjIvJr2Ay20XcaLo1-WnHQEviMUI2xUeYPYTG-2pCiv9x8Q8i96G_</recordid><startdate>19830125</startdate><enddate>19830125</enddate><creator>Kayser, R H</creator><creator>Bounds, P L</creator><creator>Bevins, C L</creator><creator>Pollack, R M</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19830125</creationdate><title>Affinity alkylation of bacterial delta 5-3-ketosteroid isomerase. 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Each of the steroid-bound peptides of the 4 beta-inhibited enzyme was subjected to further proteolytic digestion and the site of steroid attachment was found to be Asp-38 in each of the inactivation products. These results are interpreted to indicate that "backwards binding" is an important feature of the binding of steroids to delta 5-3-ketosteroid isomerase.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>6822514</pmid><doi>10.1016/S0021-9258(18)33137-5</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1983-01, Vol.258 (2), p.909-915 |
| issn | 0021-9258 1083-351X |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Amino Acids - analysis Androstenes - pharmacology Binding Sites Chromatography, High Pressure Liquid Estrenes - pharmacology Isomerases - antagonists & inhibitors Peptide Fragments - analysis Steroid Isomerases - antagonists & inhibitors Time Factors Trypsin - metabolism |
| title | Affinity alkylation of bacterial delta 5-3-ketosteroid isomerase. Identification of the amino acid modified by steroidal 17 beta-oxiranes |
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