In silico analysis of crustacean hyperglycemic hormone family G protein-coupled receptor candidates

Mihika T Kozma; Jorge L Pérez-Moreno; Neha S Gandhi; Luisanna Hernandez Jeppesen; David S Durica; Tomer Ventura; Donald L Mykles

doi:10.3389/fendo.2023.1322800

In silico analysis of crustacean hyperglycemic hormone family G protein-coupled receptor candidates

Front Endocrinol (Lausanne). 2024 Jan 9:14:1322800. doi: 10.3389/fendo.2023.1322800. eCollection 2023.

Authors

Mihika T Kozma^#¹, Jorge L Pérez-Moreno^#¹, Neha S Gandhi^{2

3}, Luisanna Hernandez Jeppesen¹, David S Durica⁴, Tomer Ventura⁵, Donald L Mykles^{1

6}

Affiliations

¹ Department of Biology, Colorado State University, Fort Collins, CO, United States.
² Department of Computer Science and Engineering, Manipal Institute of Technology, Manipal Academy of Higher Education, Manipal, Karnataka, India.
³ School of Chemistry and Physics, Queensland University of Technology, Brisbane, QLD, Australia.
⁴ Department of Biology, University of Oklahoma, Norman, OK, United States.
⁵ Centre for BioInnovation and School of Science, Technology and Engineering, University of the Sunshine Coast, Sippy Downs, QLD, Australia.
⁶ Coastal and Marine Sciences Institute, University of California-Davis Bodega Marine Laboratory, Bodega Bay, CA, United States.

^# Contributed equally.

Abstract

Ecdysteroid molting hormone synthesis is directed by a pair of molting glands or Y-organs (YOs), and this synthesis is inhibited by molt-inhibiting hormone (MIH). MIH is a member of the crustacean hyperglycemic hormone (CHH) neuropeptide superfamily, which includes CHH and insect ion transport peptide (ITP). It is hypothesized that the MIH receptor is a Class A (Rhodopsin-like) G protein-coupled receptor (GPCR). The YO of the blackback land crab, Gecarcinus lateralis, expresses 49 Class A GPCRs, three of which (Gl-CHHR-A9, -A10, and -A12) were provisionally assigned as CHH-like receptors. CrusTome, a transcriptome database assembled from 189 crustaceans and 12 ecdysozoan outgroups, was used to deorphanize candidate MIH/CHH GPCRs, relying on sequence homology to three functionally characterized ITP receptors (BNGR-A2, BNGR-A24, and BNGR-A34) in the silk moth, Bombyx mori. Phylogenetic analysis and multiple sequence alignments across major taxonomic groups revealed extensive expansion and diversification of crustacean A2, A24, and A34 receptors, designated CHH Family Receptor Candidates (CFRCs). The A2 clade was divided into three subclades; A24 clade was divided into five subclades; and A34 was divided into six subclades. The subclades were distinguished by conserved motifs in extracellular loop (ECL) 2 and ECL3 in the ligand-binding region. Eleven of the 14 subclades occurred in decapod crustaceans. In G. lateralis, seven CFRC sequences, designated Gl-CFRC-A2α1, -A24α, -A24β1, -A24β2, -A34α2, -A34β1, and -A34β2, were identified; the three A34 sequences corresponded to Gl-GPCR-A12, -A9, and A10, respectively. ECL2 in all the CFRC sequences had a two-stranded β-sheet structure similar to human Class A GPCRs, whereas the ECL2 of decapod CFRC-A34β1/β2 had an additional two-stranded β-sheet. We hypothesize that this second β-sheet on ECL2 plays a role in MIH/CHH binding and activation, which will be investigated further with functional assays.

Keywords: CrusTome; G protein-coupled receptor; Y-organ; crustacean hyperglycemic hormone; ecdysteroid; molt-inhibiting hormone; molting; neuropeptide.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Arthropod Proteins*
Benzeneacetamides*
Humans
Invertebrate Hormones*
Nerve Tissue Proteins* / genetics
Nerve Tissue Proteins* / metabolism
Phylogeny
Piperidones*
Receptors, G-Protein-Coupled* / chemistry
Receptors, G-Protein-Coupled* / genetics

Substances

hyperglycemic hormone, crustacean
antineoplaston A10
Nerve Tissue Proteins
Receptors, G-Protein-Coupled
Invertebrate Hormones
Piperidones
Benzeneacetamides
Arthropod Proteins

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was supported by grants from the National Science Foundation to DM (IOS-1922701) and DD (IOS-1922755).