Cement proteins of the acorn barnacle, Megabalanus rosa
Cement proteins of the acorn barnacle, Megabalanus rosa
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DOI：
10.2307/1543033
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发表时间：
1996-06-01
影响因子：
1.6
通讯作者：
Maruyama, T
中科院分区：
文献类型：
--
作者：
Kamino, K;Odo, S;Maruyama, T
由藤壶分泌的蛋白质粘固剂的组分由于其不溶性而尚未被研究。我们溶解和特征的次生水泥，这是藤壶是从底层分离时产生的蛋白质，在Megabalanus rosa。根据水泥在甲酸水溶液中的溶解度，将水泥分级为可溶性级分SF 1（21%）、还原后可溶性级分SF 2（37%）和还原后不溶性级分IF（42%）。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，SF 1和SF 2分别含有3条多肽（SF 1 -60 k，-57 k，-47 k）和1条多肽（SF 2 -60 k）。这些多肽的氨基酸组成相似，它们的N-末端氨基酸序列相同。这些多肽具有不寻常的氨基酸组成，富含Ser、Thr、Ala和Gly，就像海洋多毛类动物Phragmatopoma californica的管粘固剂。用溴化氰裂解后溶解在甲酸水溶液中的IF通过SDS-PAGE显示含有8个片段肽（CB-肽）。结果表明，藤壶的次级粘固物中的CB-肽与附着在基质上的初级粘固物中的CB-肽具有相同的SDS-PAGE图谱，表明藤壶的次级粘固物中至少含有两种蛋白质：SF_1和SF_2中的高羟基化蛋白和IF中的不溶性蛋白。此外，免疫印迹分析，使用多克隆抗体对CB-肽之一，从二级水泥，也交叉反应的CNBr片段肽的初级水泥。这些结果表明，初级和次级骨水泥的蛋白质组成相似。
Components of the proteinaceous cement secreted by barnacles have yet to be studied because of their insolubility. We solubilized and characterized the proteins of secondary cement, which is produced when the barnacle is detached from the substratum, in Megabalanus rosa. The cement was fractionated, according to its solubility in aqueous formic acid, into a soluble fraction, SF1 (21%); a fraction soluble after reduction, SF2 (37%); and a fraction insoluble after reduction, IF (42%). Analysis of the SF1 and SF2 by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that they contained three polypeptides (SF1-60 k, -57 k, -47 k) and one polypeptide (SF2-60 k), respectively. The amino acid compositions of these polypeptides were similar and their N-terminal amino acid sequences were identical. These polypeptides had an unusual amino acid composition, rich in Ser, Thr, Ala, and Gly, like the tube cement of a marine polychaete, Phragmatopoma californica. The IF, solubilized in aqueous formic acid after cleavage with cyanogen bromide, was shown by SDS-PAGE to contain eight fragment peptides (CB-peptides). N-terminal amino acid sequences of the CB-peptides were also determined.We conclude that the barnacle cement is composed of at least two types of protein: highly hydroxylated protein in the SF1 and SF2 and insoluble protein in the IF.The SDS-PAGE pattern of CB-peptides from the secondary cement was identical to that of the primary cement produced while the barnacle is attached to a substratum. In addition, immunoblot analysis, using a polyclonal antibody against one of the CB-peptides from the secondary cement, also cross-reacted with a CNBr-fragment peptide of the primary cement. These results indicate that the primary and secondary cements are similar in protein composition.