Department of Biochemistry, Michigan State University, 48824, East Lansing, MI, USA.
Planta. 1989 Sep;179(2):148-55. doi: 10.1007/BF00393684.
Antibodies were prepared against ratliver connexin (27-kDa polypeptide subunit of cell gap junctions found between contacting animal cells) and a putative soybean (Glycine max (L.) Merr.) connexin (29-kDa polypeptide) previously isolated from cultured soybean root cells (SB-1 cell line). The antibodies were utilized to examine the intracellular localization of soybean connexin in these cultured soybean cells and to probe for the presence of a soybean-type connexin in petals, fruits, and leaves from a variety of plants. As judged by specific reactivity on immunoblots, both antibodies against the 27-kDa polypeptide (ratliver connexin) and against the 29-kDa polypeptide (operationally termed soybean connexin) were utilized to demonstrate immunological relatedness of the 27-kDa (rat liver) and the 29-kDa (soybean) polypeptide. Immunofluorescent localization of the putative soybean connexin in cultured soybean cells utilizing these probes demonstrated a peripherally localized punctate pattern of labeling at areas of contact between cells. Use of antibody to the soybean connexin as a probe on immunoblots of extracts from petals, fruits and leaves demonstrated that the soybean-type connexin is present in a large number of different plants.
针对大鼠肝连接蛋白(细胞间隙连接中发现的接触动物细胞之间的 27kDa 多肽亚基)和先前从培养的大豆根细胞(SB-1 细胞系)中分离出的一种假定的大豆连接蛋白(29kDa 多肽)制备了抗体。这些抗体被用于研究培养的大豆细胞中大豆连接蛋白的细胞内定位,并探测各种植物的花瓣、果实和叶片中是否存在大豆型连接蛋白。通过免疫印迹上的特异性反应判断,针对 27kDa 多肽(大鼠肝连接蛋白)和针对 29kDa 多肽(操作上称为大豆连接蛋白)的两种抗体都被用于证明 27kDa(大鼠肝)和 29kDa(大豆)多肽之间的免疫学相关性。利用这些探针在培养的大豆细胞中对假定的大豆连接蛋白进行免疫荧光定位,显示出细胞接触部位的周边点状标记模式。使用大豆连接蛋白抗体作为探针在花瓣、果实和叶片提取物的免疫印迹上进行检测,表明大豆型连接蛋白存在于大量不同的植物中。