Subramanian G, Takemoto L
Division of Biology, Kansas State University, Manhattan 66506.
Invest Ophthalmol Vis Sci. 1991 Aug;32(9):2588-92.
Peptide antisera made against the putative MP18 sequence recognizes an 18-kilodalton protein on western blots of bovine lens membrane. These peptide antisera were used to investigate possible age-related changes in the MP18 molecule in membrane fractions from newborn and adult bovine lenses. Western blot analysis of membrane fractions with peptide antisera to sequences 45-50 (anti-45), 113-129 (anti-113), and C-terminus 169-173 (anti-Ct) showed similar patterns. All three antisera reacted more strongly with MP18 from membrane prepared from newborn lenses than with corresponding preparations from the adult lens. Binding ratios of anti-45 to anti-Ct showed a statistically significant difference between cortex and nucleus of newborn lenses (P less than 0.02) and between cortex of newborn and cortex of adult lenses (P less than 0.001). Differences in binding ratios of anti-113 to anti-Ct were the most striking, with anti-113 unable to detect MP18 in membrane preparations from adult lenses. Collectively, these results confirmed the putative sequence of MP18 and demonstrated covalent changes in this molecule during the process of aging in the normal bovine lens.
针对推测的MP18序列制备的肽抗血清,在牛晶状体膜的蛋白质印迹上识别出一种18千道尔顿的蛋白质。这些肽抗血清用于研究新生牛和成年牛晶状体膜组分中MP18分子可能的年龄相关变化。用针对序列45 - 50(抗-45)、113 - 129(抗-113)和C末端169 - 173(抗-Ct)的肽抗血清对膜组分进行蛋白质印迹分析,结果显示出相似的模式。所有三种抗血清与新生晶状体制备的膜中的MP18反应都比与成年晶状体相应制备物中的反应更强。抗-45与抗-Ct的结合比率在新生晶状体的皮质和核之间(P小于0.02)以及新生晶状体皮质与成年晶状体皮质之间(P小于0.001)显示出统计学上的显著差异。抗-113与抗-Ct的结合比率差异最为显著,抗-113无法检测到成年晶状体膜制备物中的MP18。总体而言,这些结果证实了MP18的推测序列,并证明了在正常牛晶状体老化过程中该分子的共价变化。