Reichmuth C, Becker S, Benz M, Debel K, Reisch D, Heimbeck G, Hofbauer A, Klagges B, Pflugfelder G O, Buchner E
Theodor-Boveri-Institut für Biowissenschaften, Lehrstuhl für Genetik, Würzburg, Germany.
Brain Res Mol Brain Res. 1995 Aug;32(1):45-54. doi: 10.1016/0169-328x(95)00058-z.
Proteins expressed specifically in neurons and transported to synaptic terminals are likely to constitute important molecular elements of nervous system function. In an effort to characterize synapse-associated proteins (SAPs) of Drosophila, we have isolated from a hybridoma library several monoclonal antibodies (MABs) that selectively stain synaptic terminals in immunohistochemical preparations. MAB nc46 binds to most but not all synaptic terminals of the Drosophila nervous system, it also recognizes a protein with homologous distribution in other dipteran flies and binds to large parts of fish CNS. In Western blots the antibody labels a Drosophila brain protein of 47 kDa and cross-reacts with brain proteins from several species including insects, fish, mouse and man. From these data we conclude that the corresponding gene has been conserved in evolution at least among diptera. Using MAB nc46 and expression cloning we have identified the 'sap47' gene coding for the 'synapse-associated protein of 47 kDa' of Drosophila melanogaster. Sequence analysis of genomic and cDNA clones reveals the intron-exon structure of the gene and characterizes the complete open reading frames of two alternatively spliced transcripts. The sap47 gene is located in 89A8-B3 on chromosome 3R and codes for two almost identical inferred polypeptides of 347 and 351 amino acids with no significant sequence homology to known proteins.
在神经元中特异性表达并转运至突触终末的蛋白质,可能构成神经系统功能的重要分子元件。为了鉴定果蝇的突触相关蛋白(SAPs),我们从杂交瘤文库中分离出几种单克隆抗体(MABs),它们在免疫组织化学制剂中能选择性地标记突触终末。单克隆抗体nc46与果蝇神经系统的大多数但并非所有突触终末结合,它还能识别在其他双翅目昆虫中具有同源分布的一种蛋白质,并与鱼类中枢神经系统的大部分区域结合。在蛋白质免疫印迹中,该抗体标记一种47 kDa的果蝇脑蛋白,并与包括昆虫、鱼类、小鼠和人类在内的几种物种的脑蛋白发生交叉反应。从这些数据我们得出结论,相应的基因至少在双翅目昆虫的进化过程中是保守的。利用单克隆抗体nc46和表达克隆技术,我们鉴定出了编码黑腹果蝇“47 kDa突触相关蛋白”的“sap47”基因。基因组和cDNA克隆的序列分析揭示了该基因的内含子-外显子结构,并确定了两种选择性剪接转录本的完整开放阅读框。sap47基因位于3R染色体的89A8-B3区域,编码两种几乎相同的推测多肽,分别为347和351个氨基酸,与已知蛋白质无明显序列同源性。