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

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, c...

Full description

Saved in:
Bibliographic Details
Published in:General and comparative endocrinology 2017-11, Vol.253, p.33-43
Main Authors: Camacho-Jiménez, Laura, Sánchez-Castrejón, Edna, Díaz, Fernando, Aguilar, Manuel B, Muñoz-Márquez, Ma Enriqueta, Ponce-Rivas, Elizabeth
Format: Article
Language:English
Subjects:
Amides - chemistry
Animals
Arthropod Proteins - genetics
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 - genetics
Invertebrate Hormones - metabolism
Nerve Tissue Proteins - genetics
Nerve Tissue Proteins - metabolism
Osmoregulation
Penaeidae - metabolism
Protein Isoforms - genetics
Protein Isoforms - metabolism
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Sequence Alignment
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0016-6480
1095-6840
DOI:10.1016/j.ygcen.2017.08.020