Cloning and expression of the recombinant crustacean hyperglycemic hormone isoform B2 (rCHH-B2) and its effects on the metabolism and osmoregulation of the Pacific white shrimp Litopenaeus vannamei.

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  • Additional Information
    • Source:
      Publisher: Academic Press Country of Publication: United States NLM ID: 0370735 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1095-6840 (Electronic) Linking ISSN: 00166480 NLM ISO Abbreviation: Gen Comp Endocrinol Subsets: MEDLINE
    • Publication Information:
      Publication: New York, NY : Academic Press
      Original Publication: New York.
    • Subject Terms:
      Osmoregulation*; Arthropod Proteins/*genetics ; Invertebrate Hormones/*genetics ; Nerve Tissue Proteins/*genetics ; Penaeidae/*metabolism; Amides/chemistry ; Animals ; Arthropod Proteins/metabolism ; Base Sequence ; Biological Assay ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Cloning, Molecular ; Computer Simulation ; Genetic Vectors/metabolism ; Hyperglycemia/metabolism ; Invertebrate Hormones/metabolism ; Nerve Tissue Proteins/metabolism ; Protein Isoforms/genetics ; Protein Isoforms/metabolism ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/metabolism ; Sequence Alignment
    • Abstract:
      Crustacean hyperglycemic hormones (CHHs) are multifunctional neuropeptides ubiquitous in crustaceans. In Litopenaeus vannamei, CHH-B2 is a CHH eyestalk isoform whose expression has been shown to vary with enviromental conditions, suggesting its relevance for ecophysiological performance of shrimp, controlling processes related to metabolism and osmo-ionic regulation. To study the involvement of CHH-B2 in these processes, we cloned and expressed a recombinant version with a free C-terminal glycine (rCHH-B2-Gly) in the methylotrophic yeast Pichia pastoris. The rCHH-B2-Gly peptide secreted to the culture medium was purified by RP-HPLC and used for in vivo glucose, triglyceride, and osmoregulation dose-response analyses with juvenile shrimp. The peptide was also amidated at the C-terminus using an α-amidating enzyme to produce rCHH-B2-amide. The shrimp showed a dose-dependent effect of rCHH-B2-Gly to hemolymph glucose and triglyceride levels, inducing maximal increases by injecting 500 and 1000pmol of hormone, respectively. Additionally, 10pmol of hormone was sufficient to reduce the hypo-osmoregulatory capacity of shrimp at 35‰. These findings suggest that CHH-B2 has regulatory roles in carbohydrate and lipid metabolism, and a potential involvement in osmoregulation of L. vannamei. Injection of 100pmol of rCHH-B2-amide increased glucose and triglyceride levels by 15 and 28%, respectively in comparison with rCHH-B2-Gly, suggesting an important role for the C-terminal amidation. Additionally, an in silico structural analysis done with the CHH-B1 and rCHH-B2-Gly peptides suggests that the C-terminal region may be relevant for the activity of the L. vannamei isoforms and explain the functional divergence from other crustacean CHH/CHH-like peptides.
      (Copyright © 2017 Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: Crustacean hyperglycemic hormone; Glucose; Lipids; Litopenaeus vannamei; Osmoregulation; Recombinant protein
    • Accession Number:
      0 (Amides)
      0 (Arthropod Proteins)
      0 (Invertebrate Hormones)
      0 (Nerve Tissue Proteins)
      0 (Protein Isoforms)
      0 (Recombinant Proteins)
      0 (hyperglycemic hormone, crustacean)
    • Publication Date:
      Date Created: 20170827 Date Completed: 20180115 Latest Revision: 20181202
    • Publication Date:
      20231215
    • Accession Number:
      10.1016/j.ygcen.2017.08.020
    • Accession Number:
      28842215