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J. Biol. Chem., Vol. 281, Issue 36, 26150-26158, September 8, 2006

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Linking Adhesive and Structural Proteins in the Attachment Plaque of Mytilus californianus^*

From the Department of Molecular, Cellular & Developmental Biology and the Department of Chemistry & Biochemistry, Marine Science Institute, University of California, Santa Barbara, California 93106

The byssal attachment of California mussels Mytilus californianus provides secure adhesion in the presence of moisture, a feat that still eludes most synthetic polymers. Matrix-assisted laser desorption ionization mass spectrometry was used to probe the footprints of byssal attachment plaques on glass cover slips for adhesive proteins. Besides the abundant mcfp-3 protein family (Zhao, H., Robertson, N. B., Jewhurst, S. A., and Waite, J. H. (2006) J. Biol. Chem. 281, 11090–11096), two new proteins, mcfp-5 and mcfp-6, with masses of 8.9 kDa and 11.6 kDa, respectively, were identified in footprints, partially characterized and completely sequenced from a cDNA library. mcfp-5 resembles mcfp-3 in its basic pI and abundant 3,4-dihydroxyphenyl-L-alanine (Dopa; 30 mol %), but is distinct in two respects: it is more homogeneous in primary sequence and is polyphosphorylated. mcfp-6 is basic and contains a small amount of Dopa (<5 mol %). In contrast to mcfp-3 and -5, tyrosine prevails at 20 mol %, and cysteine is present at 11 mol %, one-third of which remains thiolate. Given the oxidative instability of Dopa and cysteine at pH 8.2 (seawater), we tested the hypothesis that thiols serve to scavenge dopaquinones by adduct formation. Plaque footprints were hydrolyzed and screened for cysteine dopaquinone adducts using phenylboronate affinity chromatography. 5-S-Cysteinyldopa was detected at nearly 1 mol %. The results suggest that mcfp-6 may provide a cohesive link between the surface-coupling Dopa-rich proteins and the bulk of the plaque proteins.

Received for publication, May 8, 2006 , and in revised form, July 13, 2006.

^* This research was supported by in part by grants (to J. H. W.) from NASA (University Research Engineering and Technology Institute on Bio-Inspired Materials under Award No. NCC-1-02037) and the National Institutes of Health (DE015415 [GenBank] ). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) DQ351537 [GenBank] , DQ351538 [GenBank] , DQ351539 [GenBank] , and DQ444853 [GenBank] .

¹ To whom correspondence should be addressed: Marine Science Institute, University of California, Santa Barbara, CA 93106. Tel.: 805-893-2817; Fax: 805-893-7998; E-mail: waite{at}lifesci.ucsb.edu.

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