Davril M, Han K K
Int J Pept Protein Res. 1976;8(2):177-81. doi: 10.1111/j.1399-3011.1976.tb02493.x.
Highly purified elastin from porcine aorta was submitted to elastase digestion. The enzymic products were subjected to gel filtration on Sephadex G 25. The excluded fraction was then submitted to thermolysin digestion and gel filtration. The excluded fraction was submitted to partial acid hydrolysis and gel filtration. Several sub-fractions were obtained. The F3 subfraction containing cross-linking agents (desmosines and lysinonorleucine) was finally subjected to ion exchange chromatography. A highly enriched peptide fraction containing lysinomorleucine was obtained and then purified by preparative electrophoresis. The ratio of enrichment passed from 2 residues of lysinonorleucine (expressed as lysine) from starting material (elastin) to 178 out of 1,000 residues in the final step. In this peptide fraction, if we express in molar ratio and consider the amount of lysinonorleucine is one residue, the following amino-acid composition is Pro:3, Gly:1, Ala:2, LNL:1, Lys:2. No traces of desmosines are detected. The role of lysinonorleucine in bridging functions in elastin is discussed.
将从猪主动脉中高度纯化的弹性蛋白进行弹性蛋白酶消化。酶解产物在葡聚糖凝胶G 25上进行凝胶过滤。然后将排阻级分进行嗜热菌蛋白酶消化和凝胶过滤。再将排阻级分进行部分酸水解和凝胶过滤。得到了几个亚级分。最终对含有交联剂（锁链素和赖氨酰正亮氨酸）的F3亚级分进行离子交换色谱法。得到了一个高度富集的含有赖氨酰正亮氨酸的肽级分，然后通过制备电泳进行纯化。富集比从起始材料（弹性蛋白）中每1000个残基中有2个赖氨酰正亮氨酸残基（以赖氨酸表示）提高到最终步骤中的每1000个残基中有178个。在这个肽级分中，如果以摩尔比表示并考虑赖氨酰正亮氨酸的量为一个残基，其氨基酸组成为：脯氨酸：3、甘氨酸：1、丙氨酸：2、赖氨酰正亮氨酸：1、赖氨酸：2。未检测到锁链素的痕迹。文中讨论了赖氨酰正亮氨酸在弹性蛋白桥接功能中的作用。
