Li Wei, Yu Zu-Xi, Kotin Robert M
Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
J Histochem Cytochem. 2005 Aug;53(8):1003-9. doi: 10.1369/jhc.4A6568.2005. Epub 2005 May 6.
Protein kinase X (PrKX), karyotypically located on the human X chromosome, is a type I cAMP-dependent protein kinase. Although a specific role for PrKX has not yet been defined, PrKX gene expression in mouse and human tissues has been profiled only by in situ hybridization and Northern blot analyses and not by protein expression. To determine more precisely the PrKX protein levels, we developed specific anti-PrKX antibodies and examined gestationally staged mouse embryo sections by immunohistochemistry. These results showed that PrKX is ubiquitously distributed and highly expressed in murine central nervous system and heart tissues in early developmental stages and in most organs at later stages but was not detected in either connective tissues or bone. Using Western blots to detect PrKX, total protein extracts from eight different adult or fetal human tissues including brain, heart, kidney, liver, lung, pancreas, spleen, and thymus were analyzed. Although PrKX protein was present in each of the tissues tested, the protein levels varied depending on tissue type and developmental stage. Very low protein levels were found in heart tissues from a 5-month-old fetus and from an adult, whereas PrKX proteins were more abundant in fetal brain, kidney, and liver tissues compared with adult samples of the same tissue type.
蛋白激酶X(PrKX),其染色体定位在人类X染色体上,是一种I型环磷酸腺苷(cAMP)依赖性蛋白激酶。尽管PrKX的具体作用尚未明确,但小鼠和人类组织中PrKX基因的表达仅通过原位杂交和Northern印迹分析进行了研究,而未对蛋白质表达进行研究。为了更精确地确定PrKX蛋白水平,我们制备了特异性抗PrKX抗体,并通过免疫组织化学方法检测了处于不同妊娠阶段的小鼠胚胎切片。这些结果表明,PrKX在发育早期在小鼠中枢神经系统和心脏组织中广泛分布且高表达,在发育后期在大多数器官中也有表达,但在结缔组织或骨骼中均未检测到。使用蛋白质印迹法检测PrKX,对来自包括脑、心脏、肾脏、肝脏、肺、胰腺、脾脏和胸腺在内的八种不同成人或胎儿人类组织的总蛋白提取物进行了分析。尽管在每个测试组织中都存在PrKX蛋白,但蛋白水平因组织类型和发育阶段而异。在5个月大胎儿和成人的心脏组织中发现PrKX蛋白水平非常低,而与相同组织类型的成人样本相比,胎儿脑、肾脏和肝脏组织中的PrKX蛋白更为丰富。