Noma A, Takahashi T, Yamada K, Wada T
Atherosclerosis. 1979 May;33(1):29-39. doi: 10.1016/0021-9150(79)90195-3.
Elastin was extracted from human aortic plaque and adjacent grossly normal intima by the following methods: (1) 0.1 N NaOH at 100 degrees C, (2) hot NaOH and 0.2 M EDTA, (3) 5 M guanidine--HCl and collagenase, (4) guanidine--collagenase and dithioerythritol--urea--sodium dodecyl sulfate, (5) guanidine--collagenase and EDTA, (6) 10% NaCl and collagenase, and (7) NaCl--collagenase and EDTA. All elastin samples contained small amounts of carbohydrate and hydroxyproline. The lipid content of non-plaque intimal elastin samples was small (2--3%), whereas it increased to 4--6% in plaque intima. The lipid composition of elastin preparations varied significantly with the extraction procedure. Elastin from plaque intima contained significantly more cholesterol (50--60%) and less triglyceride and phospholipid than elastin of non-plaque intima (30--50% cholesterol). The contents of free and esterified cholesterol were comparable in all preparations. The main phospholipid in all samples was sphingomyelin, which comprised between 50 and 80% of the total phospholipid. Compared with NaOH-purified elastin, the other elastin samples were characterized by an increased phosphatidyl--choline content, while they all contained an almost equal amount of phosphatidylethanolamine. In elastin samples from plaque intima, the polar amino acids were increased, whereas cross-linking amino acids were decreased. The polarity and hydroxyproline content of elastin samples were slightly decreased after treatment with EDTA or dithioerythritol--urea--sodium dodecyl sulfate.
(1) 在100℃下用0.1N NaOH；(2) 热NaOH和0.2M EDTA；(3) 5M 胍-HCl和胶原酶；(4) 胍-胶原酶和二硫赤藓糖醇-尿素-十二烷基硫酸钠；(5) 胍-胶原酶和EDTA；(6) 10% NaCl和胶原酶；(⑦) NaCl-胶原酶和EDTA。所有弹性蛋白样品均含有少量碳水化合物和羟脯氨酸。非斑块内膜弹性蛋白样品的脂质含量较低(2%-3%)，而在斑块内膜中脂质含量增至4%-6%。弹性蛋白制剂的脂质组成随提取方法的不同而有显著差异。与非斑块内膜弹性蛋白(胆固醇含量为30%-50%)相比，斑块内膜弹性蛋白含有的胆固醇显著更多(50%-60%)，甘油三酯和磷脂更少。所有制剂中游离胆固醇和酯化胆固醇的含量相当。所有样品中的主要磷脂是鞘磷脂，占总磷脂的50%-80%。与NaOH纯化的弹性蛋白相比，其他弹性蛋白样品的特征是磷脂酰胆碱含量增加，而它们所含的磷脂酰乙醇胺量几乎相等。在斑块内膜的弹性蛋白样品中，极性氨基酸增加，而交联氨基酸减少。用EDTA或二硫赤藓糖醇-尿素-十二烷基硫酸钠处理后，弹性蛋白样品的极性和羟脯氨酸含量略有降低。
