Electronic spectroscopy of cobalt angiotensin converting enzyme and its inhibitor complexes.
Electronic spectroscopy of cobalt angiotensin converting enzyme and its inhibitor complexes.
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钴血管紧张素转换酶及其抑制剂复合物的电子光谱。
DOI：
10.1021/bi00397a014
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Riordan,JF
中科院分区：
文献类型：
--
作者：
Bicknell,R;Holmquist,B;Lee,FS;Martin,MT;Riordan,JF
材料与方法从幼兔肺（Pel-Freez Biologicals，Inc.）通过如前所述在Sepharose-28-CA-Phe-Gly上进行亲和层析（Pantoliano等，1984年）。亲和配体通过下述改进的程序制备，其基于Bilncher和Bilnning（1986）的方法。氯化钴为“SpecPure”级，得自约翰逊万丰有限公司，英格兰如前所述制备和测定FaFGG和FaGLA（Bünning等人，1983年）。2巯甲丙脯酸和3-巯基丙酰-精氨酸是SJ Lucania博士的礼物，Squibb医学研究所，普林斯顿，新泽西州，依那普利拉是Dr. AA Patchett，Merck，Sharp & Dohme Research Laboratories，Division of Merck and Co.，Inc.，新泽西州拉威。在早期的研究中制备了其他抑制剂（Holmquist和Vallee，1979）。CA-Phe-Gly的合成。D-苯基乳酸苄酯的制备方法为：将D-苯基乳酸的苄醇悬浮液用无水HCl气体饱和，澄清后的溶液静置16 h，加入二氯甲烷，用饱和NaHCO 3萃取，MgSO 4干燥，真空蒸馏。收集酯，为澄清油状物：bp 185 ℃（2.5 mmHg）; IR（Nujol）3480、1780、1490和1450 cm-1。向搅拌的酯溶液中lg，20 mmol）和吡啶（20 mL）。在-8 ℃下，在30分钟的时间内，将8.5 g（30 mmol）三氟甲磺酸酐加入到30 mL CH 2Cl 2中。1：1乙酸乙酯/环己烷洗脱，上样至2X 14 cm硅胶柱（Baker快速色谱级），用1：4乙酸乙酯/环己烷洗脱。产物用前部洗脱并蒸发成油状物（6.5g）。将溶于20 mL CH 2Cl 2中的油状物（4.95 g）逐滴加入到20 mL CH 2Cl 2溶液中。
Materials and MethodsMaterials. ACE was purified from young rabbit lungs (Pel-Freez Biologicals, Inc.) by affinity chromatography on Sepharose-28-CA-Phe-Gly as previously described (Pantoliano et al., 1984). The affinity ligand was prepared by the improved procedure described below, based on the method of Escher and Bilnning (1986). Cobalt chloride was of “SpecPure” grade from Johnson Matthey Co. Ltd., England. FaFGG and FaGLA were prepared and assayed as previously described (Bünning et al., 1983). 2 Captopril and 3-mercaptopropanoyl-Arg were gifts of Dr. SJ Lucania, Squibb Institute for Medical Research, Princeton, NJ, and Enalaprilat was a gift ofDr. AA Patchett, Merck, Sharp & Dohme Research Laboratories, Division of Merck and Co., Inc., Rahway, NJ. Other inhibitors were prepared in an earlier study (Holmquist & Vallee, 1979). Synthesis of CA-Phe-Gly. D-Phenyllactate benzyl ester was prepared by saturating a benzyl alcohol suspension of D-phenyllactic acid with dry HC1 gas, allowing the cleared so-lution tostand 16 h, adding methylene chloride, extracting with saturated NaHC03, drying over MgS04, and distilling under vacuum. The ester was collected as a clear oil: bp 185 C (2.5 mmHg); IR (Nujol) 3480, 1780, 1490, and 1450 cm" 1. To a stirred solution of the ester (5.1 g, 20 mmol) and pyridine (1.6 mL) in 30 mL of CH2C12 at-8 C was added 8.5 g (30 mmol) of trifluoromethanesulfonic acid anhydride over a pe-riod of 30 min. The oil obtained upon evaporation of theCH2C12 under vacuum was taken up in 1: 1 ethyl acetate/cy-clohexane and applied to a 2X 14 cm column of silica gel (Baker flash chromatography grade) and eluted with 1: 4 ethyl acetate/cyclohexane. The product eluted with the front and was evaporated to an oil (6.5 g). The oil (4.95 g) dissolved in 20 mL of CH2C12 was added dropwise to a solution of