Clarke Adam W, Arnspang Eva C, Mithieux Suzanne M, Korkmaz Emine, Braet Filip, Weiss Anthony S
School of Molecular and Microbial Biosciences G08, University of Sydney, New South Wales 2006, Australia.
Biochemistry. 2006 Aug 22;45(33):9989-96. doi: 10.1021/bi0610092.
Tropoelastin, the precursor of elastin, undergoes a rapid monomer to multimer association in an inverse temperature transition. This association culminates in the rapid formation of stable, optically distinct droplets of tropoelastin. Light scattering and microscope measurements reveal that these droplets are 2-6 microm in diameter. Scanning electron microscopy confirms that the droplets are spherical. Three-dimensional confocal image stacks based on the autofluorescence of tropoelastin reveal that droplets are loaded with hydrated tropoelastin. Droplets are viable intermediates in synthetic elastin macroassembly. Dense clusters of aggregated droplets and partially formed fibers develop when droplets are incubated in the presence of a lysyl oxidase. Lysine-reacting chemical and enzyme-assisted cross-linking conditions generate cross-linked beads due to interactions between multiple, surface-exposed lysine epsilon-amino groups. Droplets represent an efficient mechanism for the bolus delivery during elastogenesis of quantized packages of preaccreted tropoelastin.
原弹性蛋白是弹性蛋白的前体，在逆温度转变过程中经历从单体到多聚体的快速缔合。这种缔合最终导致原弹性蛋白快速形成稳定的、光学上不同的液滴。光散射和显微镜测量表明，这些液滴直径为2 - 6微米。扫描电子显微镜证实这些液滴是球形的。基于原弹性蛋白自身荧光的三维共聚焦图像堆栈显示，液滴中充满了水合原弹性蛋白。液滴是合成弹性蛋白大分子组装过程中的可行中间体。当在赖氨酰氧化酶存在的情况下孵育液滴时，会形成密集的聚集液滴簇和部分形成的纤维。赖氨酸反应性化学和酶辅助交联条件由于多个表面暴露的赖氨酸ε - 氨基之间的相互作用而产生交联珠。液滴代表了在弹性蛋白生成过程中用于定量预积聚原弹性蛋白包的团块递送的有效机制。
