利用原肌球蛋白tropomyosin探討魚類親源演化.docVIP

九孔螺(Haliotis diversicolor)肌肉原肌球蛋白之萃取與分析尤御丞*、黃銘志國立台南大學環境生態學院生物科技研究所 九孔螺(Haliotis diversicolor)是水生腹足類軟體動物，屬於鮑螺科、鮑螺屬。主要分布於日本房總半島到九州、朝鮮半島、中國南部沿海、香港及臺灣等暖海流域。哺乳動物及魚類肌肉皆具有明顯的絲狀結構，而九孔螺的肌肉結構柔軟有彈性且纖維不明顯，故對其肌肉結構及組成有相當大的興趣。 原肌球蛋白(tropomyosin, TM)廣泛存在於各種細胞中，在肌肉細胞中能藉由與肌動蛋白(actin)、肌凝蛋白(myosin)之協調促進肌肉之調節與收縮。有別於肌動蛋白與肌凝蛋白的高分子量，肌肉中的TM以284個胺基酸為主，分子量約33kDa。由於其保守性較高且較易研究，固本次研究以TM為研究對象，利用超高速離心、等電點沉澱、硫銨分劃及離子交換樹脂等方式找出精製九孔螺TM蛋白質的條件，並以一次電泳確認蛋白質純化結果。 關鍵字：原肌球蛋白、九孔螺、超高速離心 E-mail：roro900363@ Assay and purification of skeletal muscle tropomyosin from small abalone (Haliotis diversicolor) Yu-Cheng You*, Ming-Chih Huang Department of Biological Sciences and Technology, National University of Tainan Small abalone (Haliotis diversicolor) is a species of marine gastropod mollusk in the family Haliotidae and the genus Haliotis. It has widespread distribution, include coastal waters of Boso Peninsula, Kyushu, Korean Peninsula, south China and Taiwan. It is an important organism for fishery in Taiwan. Tropomyosin (TM) is widely distributed in all cell types associated with actin as a fibrous molecule composed of two α-helical chains arranged as a coiled-coil. Molecule mass of TM in the muscle cell is 33 kDa and containing 284 amino acids, it can regulate muscle contraction by interaction with actin and myosin. The perpous of this study is to confirm the method of purification of TM. We used ultracentrifugation, isoelectric point, and salting out with ammonium sulfate to extracting the crude TM from small abalone skeletal muscle. Then we purified TM by column chromatography and detected its characteristics by electrophoresis gel analysis. We hope this study can be useful for protein structure study. Keywords: tropomyosin, small abalone, ultracentrifugation E-mail：roro900363@