| New aspects concerning to the characterization and the relationship with the immune response in vivo of the spiny lobster nitric oxide synthase. | |
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MedLine Citation:
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PMID: 21971212 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Nitric oxide (NO) is a short-lived radical generated by nitric oxide synthases (NOS). NO is involved in a variety of functions in invertebrates, including host defense. In a previous study, we isolated and sequenced for the first time the NOS gene from hemocytes of Panulirus argus, demonstrating the inducibility of this enzyme by lipopolysaccharide (LPS) in vitro. In the present work, lobster hemocytes and gills exposed to Escherichia coli O55:B5 LPS showed an increase in both NOS activity and NOS gene expression in vivo. This response was dose and time dependent. The 3D NOS structure was predicted by comparative modeling showing the oxygenase and reductase domains. These domains contain the conserved binding motifs of NOS already found in a variety of organisms. The 3D structure prediction analysis allowed the selection of a fragment of 666bp that was cloned and subsequently expressed in E. coli BL21, in which a recombinant product of around 31KDa was obtained. Hyperimmune serum obtained from immunized rabbits was tested and employed to specifically detect the recombinant polypeptide or the endogenous NOS from lobster hemocytes by western blot and immunofluorescence. This study contributes to enlarge the existing knowledge related to NOS structure and NOS participation in the immune response in lobsters. The evaluation of an antibody capable to recognize NOS from lobsters constitutes a novel and interesting tool for the implementation of further studies on NOS functions in crustaceans. |
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Authors:
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Tania Rodríguez-Ramos; Yamila Carpio; Laida Ramos; Tirso Pons; Omar Farnós; Concepción Iglesias; Iván Sánchez de Melo; Yassel Ramos; Carlos Pendón; Mario Pablo Estrada; Jorge Bolívar |
Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-9-28 |
Journal Detail:
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Title: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society Volume: - ISSN: 1089-8611 ISO Abbreviation: - Publication Date: 2011 Sep |
Date Detail:
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Created Date: 2011-10-5 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9709307 Medline TA: Nitric Oxide Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011 Elsevier Inc. All rights reserved. |
Affiliation:
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Centro de Investigaciones Marinas, Universidad de La Habana (CIM-UH), Cuba. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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