Mucins and molluscan calcification. Molecular characterization of mucoperlin, a novel mucin-like protein from the nacreous shell layer of the fan mussel Pinna nobilis (Bivalvia, pteriomorphia).
A cDNA expression library constructed from mantle tissue mRNA of the Mediterranean fan mussel Pinna nobilis was screened with antibodies raised against the acetic acid-soluble shell matrix of the same species. This resulted in the isolation of a 2138-base pair cDNA, containing 13 tandem repeats of 93 base pairs. The deduced protein has a molecular mass of 66.7 kDa and a isoelectric point of 4.8. This protein, which is enriched in serine and proline residues, was overexpressed, purified, and used for producing polyclonal antibodies. Immunological in situ and in vitro tests showed that the protein is localized in the nacreous aragonitic layer of P. nobilis, but not in the calcitic prisms. Because this protein of the nacre of P. nobilis exhibits some mucin-like characteristics, we propose the name mucoperlin. This is the first paper reporting the cloning of a molluscan mucin and the first molecular evidence for the involvement of a mucin in molluscan calcification. This finding corroborates our previous hypothesis that some of the proteinaceous constituents of the molluscan shell matrix would derive from mucins, common to many metazoan lineages of the late Precambrian (Marin, F., Smith, M., Isa, Y., Muyzer, G. and Westbroek, P. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 1554-1559). The adaptation of an ancestral mucin to a new function, the regulation of the mineralization process, may be one of the molecular events, among others, that would explain the simultaneous emergence of organized calcification in many metazoan lineages during the Cambrian explosion.
Marin
F
F
Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. f.marin@chem.leidenuniv.nl
Corstjens
P
P
de Gaulejac
B
B
de Vrind-De Jong
E
E
Westbroek
P
P
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002113:Calcification, Physiologic; D002118:Calcium / Q000378:metabolism; D002119:Calcium Carbonate / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D006031:Glycosylation; D007150:Immunohistochemistry; D008522:Mediterranean Sea; D008969:Molecular Sequence Data; D008974:Mollusca / Q000235:genetics* / Q000378:metabolism; D009077:Mucins / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D020080:Tandem Repeat Sequences
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D009077:Mucins; D011994:Recombinant Proteins; C411849:mucoperlin; D002119:Calcium Carbonate; D002118:Calcium