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 Laurits Holmskov

3 Citations (Scopus)

Abstract

We have isolated a novel type of lectin named Arenicola marina lectin-1 (AML-1) from the lugworm A. 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 in the reduced state 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 the 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
Issue number51
Pages (from-to)42846-55
Number of pages10
ISSN0021-9258
DOIs
Publication statusPublished - 14 Dec 2012

Keywords

  • Amino Acid Sequence
  • Animals
  • Body Fluids
  • CHO Cells
  • Chitin
  • Cloning, Molecular
  • Cricetinae
  • DNA, Complementary
  • Electrophoresis, Polyacrylamide Gel
  • Erythrocytes
  • Glucosamine
  • Helminth Proteins
  • Helminths
  • Hemagglutination Inhibition Tests
  • Immunohistochemistry
  • Molecular Sequence Data
  • Peptides
  • Protein Binding
  • Rabbits
  • Recombinant Proteins
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Analysis, Protein
  • Sequence Homology, Amino Acid

Fingerprint

Dive into the research topics of 'Identification and characterization of a chitin-binding protein purified from coelomic fluid of the lugworm Arenicola marina defining a novel protein sequence family'. Together they form a unique fingerprint.

Cite this