Gould S J, Krisans S, Keller G A, Subramani S
Department of Biology, University of California, La Jolla 92093.
J Cell Biol. 1990 Jan;110(1):27-34. doi: 10.1083/jcb.110.1.27.
We have previously shown that the peroxisomal targeting signal in firefly luciferase consists of the COOH-terminal three amino acids of the protein, serine-lysine-leucine (Gould, S.J., G.A. Keller, N. Hosken, J. Wilkinson, and S. Subramani, 1989. J. Cell Biol. 108:1657-1664). Antibodies were raised against a synthetic peptide that contained this tripeptide at its COOH terminus. Immunofluorescence and immunocryoelectron microscopy revealed that the anti-peptide antibodies specifically detected peroxisomes in mammalian cells. Further characterization revealed that the antibodies were primarily directed against the COOH-terminal three amino acids of the peptide. In Western blot experiments, the antibodies recognized 15-20 rat liver peroxisomal proteins, but reacted with only a few proteins from other subcellular compartments. These results provide independent immunological evidence that the peroxisomal targeting signal identified in firefly luciferase is present in many peroxisomal proteins.
我们之前已经表明，萤火虫荧光素酶中的过氧化物酶体靶向信号由该蛋白质的羧基末端三个氨基酸组成，即丝氨酸 - 赖氨酸 - 亮氨酸（古尔德，S.J.，G.A.凯勒，N.霍斯肯，J.威尔金森，和S.苏布拉马尼，1989年。《细胞生物学杂志》108:1657 - 1664）。针对一种在其羧基末端包含此三肽的合成肽制备了抗体。免疫荧光和免疫冷冻电子显微镜显示，抗肽抗体能特异性检测哺乳动物细胞中的过氧化物酶体。进一步的表征表明，这些抗体主要针对该肽的羧基末端三个氨基酸。在蛋白质印迹实验中，这些抗体识别15 - 20种大鼠肝脏过氧化物酶体蛋白，但仅与来自其他亚细胞区室的少数蛋白质发生反应。这些结果提供了独立的免疫学证据，表明在萤火虫荧光素酶中鉴定出的过氧化物酶体靶向信号存在于许多过氧化物酶体蛋白中。
