Cement proteins of the acorn barnacle, Megabalanus rosa.

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.

[1]  J. Waite.  The DOPA Ephemera: A Recurrent Motif in Invertebrates. , 1992, The Biological bulletin.

[2]  U. Rüegg,et al.  [10] Reductive cleavage of cystine disulfides with tributylphosphine , 1977 .

[3]  T. Toyo’oka,et al.  Further studies on the reaction of amines and proteins with 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole , 1985 .

[4]  A. Rich,et al.  Spontaneous assembly of a self-complementary oligopeptide to form a stable macroscopic membrane. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Hamodrakas,et al.  Secondary structure predictions for silkmoth chorion proteins , 1982 .

[6]  J. Waite,et al.  Polyphenolic Substance of Mytilus edulis: Novel Adhesive Containing L-Dopa and Hydroxyproline. , 1981, Science.

[7]  Y. Inoue,et al.  Total immobilization of the extrinsic 33 kDa protein in spinach Photosystem II membrane preparations. Protein stoichiometry and stabilization of oxygen evolution , 1991 .

[8]  W. Dougherty.  Barnacle Adhesion: Reattachment of the Adult Barnacle Chthamalus Fragilis Darwin to Polystyrene Surfaces Followed by Centrifugational Shearing , 1990 .

[9]  Rzepecki Lm,et al.  Molecular diversity of marine glues: polyphenolic proteins from five mussel species. , 1991 .

[10]  A. Aitken,et al.  Performic Acid Oxidation , 1996 .

[11]  R. Laursen.  Reflections on the structure of mussel adhesive proteins. , 1992, Results and problems in cell differentiation.

[12]  J. Waite,et al.  The formation of mussel byssus: anatomy of a natural manufacturing process. , 1992, Results and problems in cell differentiation.

[13]  G. Walker.  The Biochemical Composition of the Cement of two Barnacle Species, Balanus Hameri and Balanus Crenatus , 1972, Journal of the Marine Biological Association of the United Kingdom.

[14]  H. Barnes,et al.  Further observations on the biochemical composition of the cement of Lepas fascicularis Ellis & Solander; electrophoretic examination of the protein moieties under various conditions , 1976 .

[15]  J. Waite,et al.  Cement precursor proteins of the reef-building polychaete Phragmatopoma californica (Fewkes). , 1992, Biochemistry.

[16]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[17]  Steven W. Taylor,et al.  trans-2,3-cis-3,4-Dihydroxyproline, a New Naturally Occurring Amino Acid, Is the Sixth Residue in the Tandemly Repeated Consensus Decapeptides of an Adhesive Protein from Mytilus edulis , 1994 .

[18]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[19]  A. Youngson,et al.  The biochemical composition of Lepas anatifera (L.) cement (Crustacea: Cirripedia) , 1975, Journal of the Marine Biological Association of the United Kingdom.

[20]  P. Y. Chou,et al.  Prediction of the secondary structure of proteins from their amino acid sequence. , 2006 .

[21]  J. Waite,et al.  Nature's underwater adhesive specialist , 1987 .