利用蛋白质组学手段鉴定新型贻贝足丝蛋白

贻贝通过足腺分泌特有的足丝并以此粘附于水下各种基质表面.贻贝足丝中富含各种粘附蛋白，其优异的水下粘附性能使其成为开发新型生物粘合剂的候选分子.厚壳贻贝足丝粘附能力强，本文采用尿素及盐酸胍抽提结合二维双向电泳技术（two-dimensional electrophoresis, 2-DE），分别对厚壳贻贝足丝纤维和足丝盘的蛋白质进行分离及染色;采用串联质谱技术结合常规搜库和表达序列标签（EST) 数据库搜索，对分离获得的蛋白质点进行鉴定，从中获得了mfp-3、mfp-6、胶原蛋白以及3种未曾报道过的新型贻贝足丝蛋白成分.上述研究为深入了解厚壳贻贝足丝蛋白的分子多样性、探讨其粘附机理以及从中筛选具有应用前景的贻贝足丝蛋白奠定了基础.

Identification of Novel Foot Proteins from Mytilus coruscus Byssus through Proteomic Analysis

Mussels adhere to a wide variety of submerged surfaces using the byssus secreted from their foot.Their byssal threads and the adhesive pads are composed of three distinct types of collagenous proteins,six types of foot proteins,and other two proteins.Mussel foot proteins are promising sources of water-resistant bio-adhesives or as medical adhesives for biotechnological applications.To completely define the protein composition of mussel byssus can be very challenging due to the low abundance and insoluble byssul proteins.In this study,we developed a proteomic approach for screening novel foot proteins from Mytilus coruscus.The foot proteins were extracted by urea acetate and acetate-guanidine hydrochloride were separated by two-dimensional electrophoresis(2-DE) and analyzed by matrix-assisted laser desorption/ ionization time of flight mass spectrometry(MALDI-TOF/TOF).The spectra from tandem mass spectrometer(MS/MS) were matched with the expressed sequence tag(EST) available in the public database databases for Mytilus.More than 35 mussel foot proteins were identified,including 3 novel full-length sequences of mussel foot proteins.