Identification and characterization of a chitin-binding protein purified from coelomic fluid of the lugworm Arenicola marina defining a novel protein sequence family
| Autor: | Jesper B. Moeller, Anders Schlosser, Anthony C. Willis, Uffe Holmskov, Grith Lykke Sørensen, Karsten Skjødt, Lars Vitved, Rikke Leth-Larsen, Alexandra Kharazova, Ida Tornøe, Nina Vitashenkova, Kit Peiter Lund, Søren Hansen |
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| Rok vydání: | 2012 |
| Předmět: |
DNA
Complementary Erythrocytes Protein family animal diseases Molecular Sequence Data Immunology Sequence alignment chemical and pharmacologic phenomena Chitin CHO Cells Biology Biochemistry law.invention Affinity chromatography law Chitin binding Sequence Analysis Protein hemic and lymphatic diseases Cricetinae Helminths Animals Amino Acid Sequence Cloning Molecular Molecular Biology Peptide sequence neoplasms chemistry.chemical_classification Glucosamine Molecular mass Sequence Homology Amino Acid Cell Biology Helminth Proteins Sequence Analysis DNA biochemical phenomena metabolism and nutrition Hemagglutination Inhibition Tests Molecular biology Immunohistochemistry Recombinant Proteins Amino acid Body Fluids chemistry Recombinant DNA bacteria Electrophoresis Polyacrylamide Gel Rabbits Peptides Sequence Alignment Protein Binding |
| Zdroj: | Vitashenkova, N, Moeller, J B, Leth-Larsen, R, Schlosser, A, Lund, K P, Tornøe, I, Vitved, L, Hansen, S, Willis, A, Kharazova, A D, Skjødt, K, Sorensen, G L & Holmskov, U 2012, ' Identification and characterization of a chitin-binding protein purified from coelomic fluid of the lugworm Arenicola marina defining a novel protein sequence family ', Journal of Biological Chemistry, vol. 287, pp. 42846-42855 . https://doi.org/10.1074/jbc.M112.420976 |
| ISSN: | 1083-351X |
| DOI: | 10.1074/jbc.M112.420976 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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