Identification and characterization of a chitin-binding protein purified from coelomic fluid of the lugworm Arenicola marina defining a novel protein sequence family

Nina Vitashenkova, Jesper Bonnet Moeller, Rikke Leth-Larsen, Anders Schlosser, Kit Peiter Lund, Ida Tornøe, Lars Vitved, Søren Hansen, Anthony Willis, Alexandra D Kharazova, Karsten Skjødt, Grith Lykke Sorensen, Uffe Holmskov

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

We have isolated a novel type of lectin named Arenicola marina lectin-1 (AML-1) from the lugworm Arenicola marina. The lectin was purified from the coelomic fluid by affinity chromatography on a GlcNAc-derivatized column and eluted with GlcNAc. On SDS-PAGE, AML-1 showed an apparent molecular mass of 27 and 31 kDa in the reduced state. The N-terminal amino acid sequences were identical in these two bands. In the unreduced state a complex band pattern was observed with bands from 35 kDa to more than 200 kDa. Two different full-length clones encoding polypeptides of 241 and 243 amino acids, respectively, were isolated from a coelomocyte cDNA library. The two clones designated AML-1a and AML-1b were 92% identical at the protein level, and represent a novel type of protein sequence family. Purified AML-1 induced agglutination of rabbit erythrocytes, which could be inhibited by N-acetylated saccharides. Recombinant AML-1b showed the same band pattern as the native protein, whereas recombinant AML-1a reduced lacked a 27-kDa band. AML-1b bound GlcNAc-derivatized columns and chitin, whereas AML-1a did not bind to these matrices. Immunohistochemical analysis revealed, that AML-1 is expressed by coelomocytes, in the nephridium and in round cells in epidermis and in eggs. Moreover, AML-1 expression was up-regulated in response to a parasitic infection. We conclude that AML-1 purified from coelomic fluid is encoded by AML-1b and represents a novel type of protein family that binds acetylated components.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume287
Pages (from-to)42846-42855
ISSN0021-9258
DOIs
Publication statusPublished - 2012

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