Worm peptidomics

Husson, Steven; Reumer, Ank; Temmerman, Liesbet; Haes, Wouter De; Schoofs, Liliane; Mertens, Inge; Baggerman, Geert

doi:10.1016/j.euprot.2014.04.005

Cited by 15 publications

(10 citation statements)

References 103 publications

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“…In silico studies and peptidomics previously identified 31 FLP and 52 NLP precursors in C. elegans (51,52), but none of the corresponding peptides show sequence similarity to deuterostomian TRHs or were able to activate TRHR-1 in cellculture cells. Given the presence of orphan protostomian TRHR orthologs (19)(20)(21), we hypothesized that an as yet unknown family of neuropeptide precursors in these phyla might encode the cognate ligand of TRHR-1, and we performed an in-depth in silico search.…”

Section: Discussionmentioning

confidence: 99%

Evolutionarily conserved TRH neuropeptide pathway regulates growth in Caenorhabditis elegans

Sinay

Mirabeau

Depuydt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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In vertebrates thyrotropin-releasing hormone (TRH) is a highly conserved neuropeptide that exerts the hormonal control of thyroidstimulating hormone (TSH) levels as well as neuromodulatory functions. However, a functional equivalent in protostomian animals remains unknown, although TRH receptors are conserved in proto-and deuterostomians. Here we identify a TRH-like neuropeptide precursor in Caenorhabditis elegans that belongs to a bilaterian family of TRH precursors. Using CRISPR/Cas9 and RNAi reverse genetics, we show that TRH-like neuropeptides, through the activation of their receptor TRHR-1, promote growth in C. elegans. TRH-like peptides from pharyngeal motor neurons are required for normal body size, and knockdown of their receptor in pharyngeal muscle cells reduces growth. Mutants deficient for TRH signaling have no defects in pharyngeal pumping or isthmus peristalsis rates, but their growth defect depends on the bacterial diet. In addition to the decrease in growth, trh-1 mutants have a reduced number of offspring. Our study suggests that TRH is an evolutionarily ancient neuropeptide, having its origin before the divergence of protostomes and deuterostomes, and may ancestrally have been involved in the control of postembryonic growth and reproduction.thyrotropin-releasing hormone | C. elegans | neuropeptide | molecular evolution | growth regulation A fter Harris's initial proposal on the hypothalamic control of pituitary secretion (1), it took almost two decades to identify the first hypophysiotropic molecule. In 1969 the groups of Schally and Guillemin isolated the tripeptide pQHP-NH 2 (2, 3), named "thyrotropin-releasing hormone" (TRH). The sequence of TRH is fully conserved across all vertebrates, indicating that strong evolutionary pressure has acted to preserve its structure (4). In all vertebrate phyla TRH is synthesized from a larger precursor protein (preproTRH) that contains five to eight copies of the TRH sequence (4). Following the explosion of genome and transcriptome sequence data, preproTRH was identified in chordate species lacking a bona fide pituitary, e.g., cephalochordates (5), and in the genomes of more ancient deuterostomes, including echinoderms (6, 7). In contrast to vertebrate (pQHP-NH 2 ) and chordate (pQSP-NH 2 ) tripeptide TRHs, most predicted echinoderm TRHs are tetrapeptidesLike vertebrate TRH, they are small peptides with an N-terminal pyroglutamate, a C-terminal amide (-NH 2 ) group, and amino acids with aromatic or cyclic side chains at the second and third positions. TRH therefore is widely distributed throughout the deuterostomian lineage of the Animal Kingdom, suggesting an ancient origin for this neuropeptide hormone.In mammals, hypothalamic TRH is the prime regulator of the set point of thyroid-stimulating hormone (TSH) synthesis and secretion by the anterior pituitary thyrotrophs (9). TSH secretion stimulates the thyroid gland to produce the thyroid hormones (THs) thyroxine (T 4 ) and triiodothyronine (T 3 ). The hypothalamus-pituitary-thyroid (HPT) axis is essentia...

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Section: Discussionmentioning

confidence: 99%

Evolutionarily conserved TRH neuropeptide pathway regulates growth in Caenorhabditis elegans

Sinay

Mirabeau

Depuydt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…This pool was aliquoted into 10 samples, each containing approximately 1 ml of biological material. Peptides were extracted using an acidified methanol extraction solvent as described previously [31]. This precipitates larger proteins, while smaller peptides remain in solution.…”

Section: Extraction Of Peptidesmentioning

confidence: 99%

“…Neuropeptidomics have revealed thousands of bioactive peptides, guiding functional studies on neuropeptide functions. Often, small invertebrate model organisms have been favored for such studies [5,[29][30][31][32]. C. elegans has emerged as a powerful model to study peptidergic modulation on a cellular and molecular level, facilitated by its completely mapped nervous system constituting 302 neurons [30,33,34].…”

Section: Introductionmentioning

confidence: 99%

A Caenorhabditis elegans Mass Spectrometric Resource for Neuropeptidomics

Bael

Zels

Boonen

et al. 2018

J. Am. Soc. Mass Spectrom.

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Neuropeptides are important signaling molecules used by nervous systems to mediate and fine-tune neuronal communication. They can function as neurotransmitters or neuromodulators in neural circuits, or they can be released as neurohormones to target distant cells and tissues. Neuropeptides are typically cleaved from larger precursor proteins by the action of proteases and can be the subject of post-translational modifications. The short, mature neuropeptide sequences often entail the only evolutionarily reasonably conserved regions in these precursor proteins. Therefore, it is particularly challenging to predict all putative bioactive peptides through in silico mining of neuropeptide precursor sequences. Peptidomics is an approach that allows de novo characterization of peptides extracted from body fluids, cells, tissues, organs, or whole-body preparations. Mass spectrometry, often combined with on-line liquid chromatography, is a hallmark technique used in peptidomics research. Here, we used an acidified methanol extraction procedure and a quadrupole-Orbitrap LC-MS/MS pipeline to analyze the neuropeptidome of Caenorhabditis elegans. We identified an unprecedented number of 203 mature neuropeptides from C. elegans whole-body extracts, including 35 peptides from known, hypothetical, as well as from completely novel neuropeptide precursor proteins that have not been predicted in silico. This set of biochemically verified peptide sequences provides the most elaborate C. elegans reference neurpeptidome so far. To exploit this resource to the fullest, we make our in-house database of known and predicted neuropeptides available to the community as a valuable resource. We are providing these collective data to help the community progress, amongst others, by supporting future differential and/or functional studies. Graphical Abstract ᅟ.

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“…WT, pamn-1, pghm-1, pgal-1, pghm-1;pamn-1, and tbh-1 mixed stage worms from 24 fully grown Petri dishes (inner diameter, 90 mm) were collected, and peptides were extracted using an acidified methanol extraction solvent as described previously (2,4,5,45,46). This extraction solvent has the advantage of precipitating larger proteins, whereas smaller peptides remain in solution.…”

Section: Extraction Of Endogenous Peptidesmentioning

confidence: 99%

Mass spectrometric evidence for neuropeptide-amidating enzymes in

Bael

Watteyne

Boonen

et al. 2018

Journal of Biological Chemistry

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Neuropeptides constitute a vast and functionally diverse family of neurochemical signaling molecules and are widely involved in the regulation of various physiological processes. The nematode is well-suited for the study of neuropeptide biochemistry and function, as neuropeptide biosynthesis enzymes are not essential for viability. This permits the study of neuropeptide biosynthesis in mutants lacking certain neuropeptide-processing enzymes. Mass spectrometry has been used to study the effects of proprotein convertase and carboxypeptidase mutations on proteolytic processing of neuropeptide precursors and on the peptidome in However, the enzymes required for the last step in the production of many bioactive peptides, the carboxyl-terminal amidation reaction, have not been characterized in this manner. Here, we describe three genes that encode homologs of neuropeptide amidation enzymes in and used tandem LC-MS to compare neuropeptides in WT animals with those in newly generated mutants for these putative amidation enzymes. We report that mutants lacking both a functional peptidylglycine α-hydroxylating monooxygenase and a peptidylglycine α-amidating monooxygenase had a severely altered neuropeptide profile and also a decreased number of offspring. Interestingly, single mutants of the amidation enzymes still expressed some fully processed amidated neuropeptides, indicating the existence of a redundant amidation mechanism in All MS data are available via ProteomeXchange with the identifier PXD008942. In summary, the key steps in neuropeptide processing in seem to be executed by redundant enzymes, and loss of these enzymes severely affects brood size, supporting the need of amidated peptides for reproduction.

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Worm peptidomics

Cited by 15 publications

References 103 publications

Evolutionarily conserved TRH neuropeptide pathway regulates growth in Caenorhabditis elegans

Evolutionarily conserved TRH neuropeptide pathway regulates growth in Caenorhabditis elegans

A Caenorhabditis elegans Mass Spectrometric Resource for Neuropeptidomics

Mass spectrometric evidence for neuropeptide-amidating enzymes in

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