Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Enrichment and Fragmentation Approaches for Enhanced Detection and Characterization of Endogenous Glycosylated Neuropeptides

ABSTRACT: Glycosylation, a common and heterogeneous post-translational modification plays a key role in altering biomolecule function and other properties. Prior to monitoring regulatory roles, basal glyconeuropeptide expression must be determined. Characterization is partially limited by the lower abundance of glycosylated neuropeptides in vivo compared to non-modified neuropeptides and thus requires effective enrichment and characterization strategies. A hydrophilic interaction liquid chromatography enrichment strategy is modified, and fragmentation schemes are evaluated to establish a workflow for future glyconeuropeptide studies to improve the understanding of neuropeptide glycosylation, as well as its distribution across the neuroendocrine system and several neuropeptide families. Product ion-triggered electron-transfer/higher-energy collision dissociation and stepped collision energy higher-energy collisional dissociation are evaluated.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive

ORGANISM(S): Callinectes Sapidus

TISSUE(S): Brain, X-organ-sinus-gland Complex, Thoracic Ganglion

SUBMITTER: Ashley Phetsanthad

LAB HEAD: Lingjun Li

PROVIDER: PXD033226 | Pride | 2022-08-19

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

HyPep: An Open-Source Software for Identification and Discovery of Neuropeptides using Sequence Homology Searching

Project description:Neuropeptides are a class of endogenous peptides that have key regulatory roles in biochemical, physiological, and behavioral processes. Mass spectrometry analyses of neuropeptides often rely on protein informatics tools for database searching and peptide identification. As neuropeptide databases are typically experimentally built and comprised of short sequences with high sequence similarity to each other, we developed a novel database searching tool, HyPep, which utilizes sequence homology searching for peptide identification. HyPep aligns database-free, de novo sequenced peptide sequences generated through PEAKS software with neuropeptide database sequences to identify neuropeptides based on an alignment score. HyPep performance was optimized using LC-MS/MS measurements of peptide extracts from various C. sapidus neuronal tissue types and compared with a commercial database searching software, PEAKS DB. HyPep identified more neuropeptides from each tissue type than PEAKS DB at 1% false discovery rate and the false match rate from both programs was 2%. In addition to identification, this report describes how HyPep can aid in the discovery of novel neuropeptides.

2023-02-02 | PXD037058 | Pride

Multifaceted Mass Spectrometric Investigation of Neuropeptide Changes in Atlantic Blue Crab, Callinectes sapidus, in Response to Low pH Stress

Project description:The decrease of pH level in the water affects animals living in aquatic habitat, such as crustaceans. The molecular mechanisms enabling these animals to survive this environmental stress remain unknown. To understand the modulatory function of neuropeptides in crustaceans when encountering drops in pH level, we developed and implemented a multifaceted mass spectrometric platform to investigate the global neuropeptide changes in response to water acidification in the Atlantic blue crab, Callinectes sapidus. Neural tissues were collected at different incubation periods to monitor dynamic changes of neuropeptides under different stress conditions occurring in the animal. Neuropeptide families were found to exhibit distinct expression patterns in different tissues and even each isoform had its specific response to the stress. Circulating fluid in the crabs (hemolymph) was also analyzed after 2-hour exposure to acidification condition, and together with results from tissue analysis, enabled the discovery of neuropeptides participating in the stress accommodation process as putative hormones. Two novel peptide sequences were detected in the hemolymph that appeared to be involved in the stress-related regulation in the crabs.

2021-09-08 | PXD013536 | Pride

Mass spectrometric evidence for neuropeptide-amidating enzymes in C. elegans

Project description: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 C. elegans is well-suited for the study of neuropeptide biochemistry and function, as neuropeptide biosynthesis enzymes are not essential for C. elegans 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 C. elegans. However, the enzymes required for the last step in the production of many bioactive peptides – the carboxyterminal amidation reaction – have not been characterized in this manner. Here, we describe three genes that encode homologs of neuropeptide amidation enzymes in C. elegans and used tandem LC-MS to compare neuropeptides in wild-type animals with those in newly generated mutants for these putative amidation enzymes. We report that mutants lacking both a functional peptidylglycine α-hydroxylating monooxygenase (PHM) and a peptidylglycine α-amidating monooxygenase (PAM) 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 C. elegans. In summary, the key steps in neuropeptide-processing in C. elegans seem to be executed by redundant enzymes, and loss of these enzymes severely affects brood size, supporting the need of amidated peptides for C. elegans reproduction.

2018-03-12 | PXD008942 | Pride

Signature-Ion Triggered Mass Spectrometry Approach Enabled Discovery of N- and O-linked Glycosylated Neuropeptides in the Crustacean Nervous System

Project description:Discovery of N-linked and O-linked glycosylation on crustacean neuropeptides.

2021-09-08 | PXD014269 | Pride

Mass spectrometry-driven discovery of neuropeptides mediating nictation behavior of nematodes

Project description:Very little is known about the genetic modulation of nictation, a host-finding strategy used by infective juveniles of many pathogenic nematodes, as well as a phoretic strategy of Caenorhabditis elegans dauers. We here use a mass spectrometry-driven approach to find neuropeptides involved in the modulation of this behavior. We identified 126 neuropeptides in infective juveniles of the commercially relevant entomopathogenic nematode Steinernema carpocapsae, 75 of which have orthologs in the model organism C. elegans. To prioritize candidates for causality testing in light of the stage-restricted nictation behavior, we developed quantitative peptidomic assays and carried out targeted proteomic measurements that allows comparing neuropeptide levels at different C. elegans life stages. Our results show that virtually all 164 quantified neuropeptides are more abundant in dauers compared to L3 juveniles. Even so, not all are relevant to nictation, and we identify the neuropeptide genes flp-7 and flp-11 as novel regulators of nictation.

2022-12-08 | PXD034629 | Pride

Deorphanization of neuropeptide GPCRs in Nematostella vectensis

Project description:The aim of this part of the wider project is to identify neuropeptide precursors, investigate cleavage sites on neuropeptide precursors and predict mature peptides in the sea anemone Nematostella vectensis. This was done to create a synthetic library of N. vectensis neuropeptides which were then used to test neuropeptide receptor candidates for activation by the different peptides.

2024-04-08 | PXD041235 | Pride

Genomics- and Peptidomics-Based Discovery of Conserved and Novel Neuropeptides in the American Cockroach

Project description:As a model hemimetabolous insect species and an invasive urban pest that is globally distributed, the American cockroach, Periplaneta americana, is of great interest in both basic and applied research. Previous studies on P. americana neuropeptide identification have been based on biochemical isolation and molecular cloning. In the present study, an integrated approach of genomics- and peptidomics-based discovery was performed for neuropeptide identification in this insect species. Using large-scale peptidomic analysis of peptide extracts from 4 different tissues (the central nervous system, corpora cardiac and corpora allata complex, midgut, and male accessory gland), 35 conserved (predicted) and a potential (novel) neuropeptides were then identified. Subsequent experiments revealed the tissue distribution, sex difference, and developmental patterns of 2 conserved neuropeptides (allatostatin B and short neuropeptide F) and a novel neuropeptide PaOGS36577. Our study shows a comprehensive neuropeptidome and detailed spatiotemporal distribution patterns, providing a solid basis for future functional studies of neuropeptides in the American cockroach.

2020-11-12 | PXD021660 | Pride

Definitive Screening Design to Improve Data-Independent MS Analysis of Neuropeptides

Project description:A definitive screening design was used to systematically optimize a DIA workflow for crustacean neuropeptide quantitation. We were able to assess several parameters for their effect on increasing quantifiable neuropeptides and predict the optimal value for these parameters.

2023-05-10 | PXD038520 | Pride

Neuropeptidomic Analysis of the Stick Insect, <i> Carausius morosus </i>

Project description:One of the most thoroughly studied insect species, with respect to locomotion behaviour, is the stick insect Carausius morosus. Although detailed information exists on premotor networks controlling walking, surprisingly little is known about neuropeptides, which are certainly involved in motor activity generation and modulation. So far, only few neuropeptides were identified from C. morosus or related stick insects. We performed a transcriptome analysis of the central nervous system to assemble and identify 65 neuropeptide and protein hormone precursors of C. morosus, including five novel putative neuropeptide precursors without clear homology to known neuropeptide precursors of other insects (Carausius neuropeptide-like precursor 1, HanSolin, PK-like1, PK-like2, RFLamide). Using Q Exactive Orbitrap and MALDI-TOF mass spectrometry, 277 peptides including 153 likely bioactive mature neuropeptides were confirmed. Peptidomics yielded a complete coverage for many of the neuropeptide propeptides and confirmed a surprisingly high number of heterozygous sequences. Few neuropeptide precursors commonly occurring in insects, including those of insect kinins and sulfakinins, could neither be found in the transcriptome data nor did peptidomics support their presence. The results of our study represent one of the most comprehensive peptidomic analyses on insects and provide the necessary input for subsequent experiments revealing neuropeptide function in greater detail.

2018-04-30 | PXD009536 | Pride

A Temporal Study of the Perturbation of Crustacean Neuropeptides Due to Severe Hypoxia Using 4-Plex Reductive Dimethylation

Project description:Hypoxia (i.e., low oxygen (O2) levels) is a common environmental challenge for several aquatic species, including fish and invertebrates. To survive or escape these conditions, these animals have developed novel biological mechanisms, some regulated by neuropeptides. By utilizing mass spectrometry, this study aims to provide a global perspective of neuropeptides in the blue crab, Callinectes sapidus, and their changes over time (0, 1, 4, and 8 hours) due to severe hypoxia (~10% O2 water saturation) stress using a 4-plex dimethyl labeling strategy to increase throughput. Using both electrospray ionization and matrix-assisted laser desorption/ionization provided complementary coverage– 88 neuropeptides were identified with only five overlapping between the two ionization techniques. Interesting trends include (1) an overall decrease in expression due to hypoxia exposure, (2) a return to basal levels after 4 or 8 hours of exposure following an initial response, (3) changes only after 4+ hours exposure, and (4) an oscillating pattern. Overall, this study boosts the power of multiplexed quantitation to understand the large-scale changes due to severe hypoxia stress over time.

2020-03-05 | PXD014688 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data