Soskel N T
Anal Biochem. 1987 Jan;160(1):98-104. doi: 10.1016/0003-2697(87)90618-x.
Quantitation of desmosine and isodesmosine, the major crosslinks in elastin, has been of interest because of their uniqueness and use as markers of that protein. Accurate measurement of these crosslinks may allow determination of elastin degradation in vivo and elastin content in tissues, obviating lengthy extraction procedures. We have developed a method of quantitating desmosine plus isodesmosine in hydrolysates of tissue and insoluble elastin using high-performance liquid chromatographic separation and absorbance detection that is rapid (21-35 min) and sensitive (accurate linearity from 100 pmol to 5 nmol). This method has been used to quantitate desmosines in elastin from bovine nuchal ligament and lung and in whole aorta from hamster. The ability to completely separate [3H]lysine from desmosine plus isodesmosine allows the method to be used to study incorporation of lysine into crosslinks in elastin.
由于弹性蛋白中的主要交联物锁链素和异锁链素具有独特性并可作为该蛋白质的标志物，因此对其进行定量分析一直备受关注。准确测量这些交联物可以确定体内弹性蛋白的降解情况以及组织中的弹性蛋白含量，从而避免冗长的提取过程。我们开发了一种方法，利用高效液相色谱分离和吸光度检测来定量组织水解产物和不溶性弹性蛋白中的锁链素加异锁链素，该方法快速（21 - 35分钟）且灵敏（在100皮摩尔至5纳摩尔范围内具有准确的线性关系）。此方法已用于定量牛颈部韧带和肺中的弹性蛋白以及仓鼠整个主动脉中的锁链素。能够将[³H]赖氨酸与锁链素加异锁链素完全分离，使得该方法可用于研究赖氨酸掺入弹性蛋白交联物的情况。
