摘要
采用透射电子显微镜观察了虾夷盘扇贝(Patinopecten yessoensis)鳃组织细胞的超微结构,发现镉盐能胁迫鳃组织中的腮丝、细胞核和线粒体产生病变.利用双向凝胶电泳(2D-PAGE)优化分离扇贝鳃组织的全蛋白,获得约800个蛋白质斑点,并筛选出37个由于镉盐胁迫而产生的差异蛋白质斑点.选用基质辅助激光解吸离子化-飞行时间质谱(MALD I-TOF MS)技术和数据库检索鉴定差异蛋白,结果发现7个与镉毒性密切相关的蛋白质,即热休克蛋白70和β-淀粉酶等上调蛋白质及原肌球蛋白、肌动蛋白和钙活化核苷酸酶1等下调蛋白质.此外,还发现转录调节子Crp/Fnr家族为低表达蛋白质,而ABC转运子为高表达蛋白质.在这些差异蛋白中,部分蛋白质适合作为连续监测流动海水中镉污染程度及评价其危害性的蛋白指示物.
The organelle ultrastructure in gill tissue was observed by transmission electron microscope（TEM） in Patinopecten yessoensis（PY）,finding the pathological changes of gill filaments,nucleus and mitochondria in the tissue exposed to CdCl2（10 mg/L）.The proteome of gill tissue in PY was perfectly separated by two-dimensional polyacrylamide gel electrophoresis（2D-PAGE）,obtaining approximately 800 protein spots,and selecting 37 differential spots intimidated with cadmium salt in the gel.In addition,these differential proteins were identified by both matrix-assisted laser desorption/ionization-time of flight mass spectrometry（MALDI-TOF MS） and database searching.The results showed that 7 differential proteins tightly connected with the cadmium toxicity were considered to be up-regulated proteins such as heat shock protein 70 and β-amylase,and down-regulated proteins such as tropomyosin,actin and calcium activated nucleotidase 1.Moreover,transcriptional regulator Crp/Fnr family showed low expression,while ABC transporter showed high expression.We suggest that these differential proteins in part have strong potentials to be utilized as protein biomarkers for monitoring the pollution level of cadmium continuously and evaluating its risk to organisms.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2010年第3期507-513,共7页
Chemical Journal of Chinese Universities
基金
国家自然科学基金(批准号:04776060)
国家“九七三”计划项目(批准号:2010CB12640)资助
关键词
蛋白质组学
虾夷盘扇贝
鳃
镉胁迫
应激蛋白
Proteomics
Patinopecten yessoensis
Gill
Cadmium stress
Stress protein
作者简介
联系人：黄河清，男，教授，博士生导师，主要从事蛋白质结构与功能及蛋白质组学研究．E—mail：hqhuang@xmu．edu．cn
