12-[(5-iodo-4-azido-2-hydroxybenzoyl)amino]dodecanoic acid: biological recognition by cholesterol esterase and acyl-CoA:cholesterol O-acyltransferase.
12-[(5-iodo-4-azido-2-hydroxybenzoyl)amino]dodecanoic acid: biological recognition by cholesterol esterase and acyl-CoA:cholesterol O-acyltransferase.
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12-[(5-碘-4-叠氮基-2-羟基苯甲酰基)氨基]十二烷酸：胆固醇酯酶和酰基辅酶A：胆固醇O-酰基转移酶的生物识别。
DOI：
10.1021/bi00458a042
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Lange,LG
中科院分区：
文献类型：
--
作者：
Kinnunen,PM;Klopf,FH;Bastiani,CA;Gelfman,CM;Lange,LG
修订稿于 1989 年 10 月 9 日收到 摘要：合成了蛋白质胆固醇和脂肪酸结合位点的潜在探针，即 12-[(5-碘-4-叠氮基-2-羟基苯甲酰基)氨基]十二烷酸酯 (IFA) 及其辅酶 A (IFA: CoA) 和胆固醇酯 (IFA: CEA)。这些放射性、光反应性脂质类似物被认为是酰基辅酶A的底物和抑制剂：胆固醇0-酰基转移酶(ACAT)和胆固醇酯酶、中性脂质结合酶，它们是调节细胞胆固醇代谢的关键要素。在黑暗中，IFA 可逆地抑制纯化的牛胰腺胆固醇酯酶对胆固醇 [14C] 油酸酯的水解，表观 K 值为 150/uM。在用紫外线光解后，IFA 对胆固醇酯酶的抑制作用变得不可逆，而油酸 (1 mM) 提供了 50% 的灭活保护。用 IFA/ 孵育均质牛胰胆固醇酯酶。 CEA 导致其水解为 IFA 和胆固醇，表明 IFA: CEA 被胆固醇酯酶识别为底物。辅酶 A 酯 (IFA: CoA) 在黑暗条件下（表观 K= 20 mM）是微粒体 ACAT 活性的可逆抑制剂，光解导致酶活性不可逆抑制，效率为 87%。 IFA: CoA 也被肝脏和主动脉微粒体 ACAT 识别为底物，从而合成 125IFA: CEA。因此，IFA 及其衍生物 IFA:CEA 和 IFA:CoA 是胆固醇酯酶和 ACAT 的抑制剂和底物。这些光亲和脂质类似物的生物识别将有助于迄今为止尚未表征的蛋白质脂质结合位点的鉴定和结构分析。
Revised Manuscript Received October 9, 1989 abstract: Potential probes of protein cholesterol and fatty acid binding sites, namely, 12-[(5-iodo-4-azido-2-hydroxybenzoyl) amino] dodecanoate (IFA) and its coenzyme A (IFA: CoA) and cholesteryl (IFA: CEA) esters, were synthesized. These radioactive, photoreactive lipid analogues were recognized as substrates and inhibitors of acyl-CoA: cholesterol 0-acyltransferase (ACAT) and cholesterol esterase, neutral lipid binding enzymes which are key elements in the regulation of cellular cholesterol metabolism. In the dark, IFA reversibly inhibited cholesteryl [14C] oleate hydrolysis by purified bovine pancreatic cholesterol esterase with an apparent K, of 150/uM. Cholesterol esterase inhibition by IFA became irreversible after photolysis with UV light and oleic acid (1 mM) provided 50% protection against inactivation. Incubation of homogeneous bovine pancreatic cholesterol esterase with IFA/. CEA resulted in its hydrolysis to IFA and cholesterol, indicating recognition of IFA: CEA as a substrate by cholesterol esterase. The coenzyme A ester, IFA: CoA, was a reversible inhibitor of microsomal ACAT activity under dark conditions (apparent K,= 20 mM), and photolysis resulted in irreversible inhibition of enzyme activity with 87% efficiency. IFA: CoA was also recognized as a substrate by both liver and aortic microsomal ACATs, with resultant synthesis of 125IFA: CEA. IFA and its derivatives, IFA: CEA and IFA: CoA, are thus inhibitors and substrates for cholesterol esterase and ACAT. Biological recognition of these photoaffinity lipid analogues will facilitate the identification and structural analysis of hithertouncharacterized protein lipid binding sites.