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作 者:刘艳[1,2] 侯付景 周君[1] 王中华[1] 李太武[4] 苏秀榕[1]
机构地区:[1]宁波大学海洋学院,宁波315211 [2]宁波职业技术学院应用化工系,宁波315800 [3]宁波绿之健药业有限公司,宁波315505 [4]宁波城市职业技术学院,宁波315100
出 处:《海洋与湖沼》2013年第4期973-979,共7页Oceanologia Et Limnologia Sinica
基 金:国家自然科学基金资助项目;"星虫铁结合蛋白基因的真核表达及富铅功能的解析";41176123号;国家自然科学基金资助项目;"星虫富集汞;铅;砷有毒重金属的分子生物学机理解读";40776075号;海洋公益性行业专项经费资助项目;"海洋污染生物效应快速监测与评价技术应用示范";201005106-2号
摘 要:以克隆获得的可口革囊星虫(Phasolosma esculenta)、枝吻纽虫(Dendrorhynchus zhejiangensis)、泥蚶(Tegillarca granosa)、缢蛏(Sinonovacula constricta)、仿刺参(Apostichopus japonicus)和白肛海地瓜(Acaudina leucoprocta)6种无脊椎动物的铁结合蛋白基因为基础,通过NCBI检索下载相应的氨基酸序列,采用SingalP程序查找信号肽,用TMHMM程序搜寻预测跨膜区,采用Clustal W程序多序列比对,构建进化树。对其编码蛋白的信号肽、跨膜区以及磷酸化位点等进行分析。结果表明,6种动物的铁结合蛋白都没有信号肽,也无跨膜区,不可能是膜上的受体或定位于膜上,均为胞外蛋白。泥蚶的ferritin有4个磷酸化位点,仿刺参ferritin有10个,枝吻纽虫有8个,白肛海地瓜有9个,可口革囊星虫和缢蛏的ferritin未搜寻到磷酸化位点。6种铁结合蛋白的疏水性有一定的差异,最大值在1.0—1.6之间,最小值在3.3—2.8之间。可口革囊星虫和仿刺参的铁结合区域特征序列完全相同,与文昌鱼、缢蛏、泥蚶、牡蛎、盘鲍有2个氨基酸差异,与人、小家鼠和叉尾有1个氨基酸差异。Based on the ferritin genes obtained in laboratory (including genes from Phascolosoma esculenta, Dendrorhynchus zhejiangensis, Tegillarca granosa, Sinonovacula constricta, Apostichopus japonicus, and Acaudina leucoprocta), other ferritin genes were found in the NCBI database, and related amino acid sequences were downloaded. The SingalP program was used to search for signal peptides. The TMHMM program was used to search for and predict the transmembrane domain. The NetPhos program was used to predict the phosphorylation sites. ProtScale was used to analyze protein hydrophobicity. Clustal W was used to make multiple sequence alignments. The phylogenetic tree was constructed. The results show that none of the ferritins had any signal peptides or transmembrane domains. None could be located on the cell membrane. The ferritin of T. granosa was predicted to have four phosphorylation sites, the ferritin of D. zhejiangensis have eight; that of , nine; and that of A. japonicus, ten. The ferritins of P. esculenta and S. constricta were predicted to have no phosphorylation sites. The hydrophobicity of ferritin showed some differences across these six species. The maximum hydrophobicity appeared at 1—1.6 and the minimum value was within the range of ?3.3— ?2.8. Multiple sequence alignment showed that ferritin had an iron ion binding region (EEREHAEKLM FQNQRGGR) and N-glycosylation site (NQSL) base sequences and that both these areas were highly conserved, indicating that the role of ferritin in iron metabolism may be significant.
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