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Neutrophil Defensins (Alpha-Defensins)

ABSTRACT: In the HPLC high-resolution ESI-MS profiles of Gingival Crevicular Fluid, four proteins, with exp. monoisotopic ion [M+H]+ at 3440.54 ± 0.06 m/z, 3369.50± 0.06 m/z, 3484.51 ± 0.06 m/z and 3707.77 ± 0.06, eluting between 24.5-26.6 minutes, were detected. The monoisotopic mass values and the manual inspection of the high-resolution MS/MS spectra of the ions at [M+5H]+5 at 689.31, 675.10, 698.11 and 742.76 m/z, and of the ions at [M+4H]+4 at 843.63, 872.39 m/z allowed to establish that the four proteins were alpha defensins 1, 2, 3 and 4.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Federica Iavarone

LAB HEAD: Federica Iavarone

PROVIDER: PXD016049 | Pride | 2020-07-09

REPOSITORIES: Pride

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Top down proteomic analysis of gingival crevicular fluid in deciduous, exfoliating and permanent teeth in children.

Iavarone Federica F Olianas Alessandra A Patini Romeo R Gallenzi Patrizia P Di Tonno Laura L Desiderio Claudia C Cabras Tiziana T Manconi Barbara B Vincenzoni Federica F Cordaro Massimo M Messana Irene I Urbani Andrea A Castagnola Massimo M

Journal of proteomics 20200703

Gingival Crevicular Fluid (GCF), a plasma-derived exudate present in the gingival crevice was collected from deciduous, exfoliating and permanent teeth from 20 children (60 samples) with the aim to characterize and quantify by a mass spectrometry based top-down proteomic approach, the peptide/proteins in the fluid and verify possible variations occurring during the exfoliating process. The results obtained confirmed the presence in GCF of α-Defensins 1-4, Thymosin β4 and Thymosin β10, as describ ...[more]

PMID: 32629195

Similar Datasets

Top Down Proteomic analysis of Gingival Crevicular Fluid in deciduous, exfoliating and permanent teeth in children

Project description:Gingival Crevicular Fluid, a plasma-derived exudate present in the gingival crevice was collected from deciduous, exfoliating and permanent teeth from 20 children (60 samples) with the aim to characterize and quantify by a mass spectrometry based top-down proteomic approach, the peptide/proteins in the fluid and verify possible variations occurring during the exfoliating process. The results obtained confirmed the presence in Gingival Crevicular Fluid of α-Defensins 1-4, Thymosin β4 and Thymosin β10, as described in previous works and revealed the presence of other interesting peptides never described before in Gingival Crevicular Fluid, such as specific fragments of α-1-antitrypsin, α-1-antichymotrypsin, Thymosin β4 and Thymosin β10 fragments, Fibrinopeptide A, Fibrinopeptide B, S100A, LVV Hemorphin-7 (hemoglobin chain β fragment), as well as some other peptides deriving from α and β subunits of hemoglobin. Statistical analysis evidenced different levels in 5 proteins/peptides in the three groups in particular with higher level in exfoliating teeth. Our study demonstrate that an in-depth analysis of a biological fluid like Gingival Crevicular Fluid, present in small amount, can provide useful information for the understanding of different biological processes like teeth eruption.

2020-07-09 | PXD016010 | Pride

Human Salivary Neutrophil Defensins (Alpha-Defensins)

Project description:In the HPLC high-resolution ESI-MS profiles of both Predominantly Antibody Deficiency syndromes subjects and healthy controls saliva samples, four proteins, with exp. monoisotopic ion [M+H]+ at 3440.54 ± 0.06 m/z, 3369.50± 0.06 m/z, 3484.51 ± 0.06 m/z and 3707.77 ± 0.06, eluting between 24.5-26.6 minutes, were detected. The monoisotopic mass values and the manual inspection of the high-resolution MS/MS spectra of the ions at [M+5H]+5 at 689.31, 675.10, 698.11 and 742.76 m/z, and of the ions at [M+4H]+4 at 843.63, 872.39 m/z allowed to establish that the four proteins were alpha defensins 1, 2, 3 and 4.

2020-01-09 | PXD012688 | Pride

Human Salivary Prolactine Inducible Protein (PIP)

Project description:In the HPLC high-resolution ESI-MS profiles of both multiple sclerosis subjects and healthy controls saliva samples three proteins, with exp. monoisotopic ion [M+H]+ at 13493.9 ± 0.2 m/z, 13696.9 ± 0.2 m/z (+203 Da, with respect to 13493.9), and 13843.1 ± 0.2 m/z (+147 Da, with respect to 13696.9) eluting between 37.8-38.2 minutes, were detected. The mass difference of 203 Da suggested that the protein with exp. monoisotopic [M+H]+ at 13696.9 ± 0.2 m/z could correspond to the N-acetylhexosamine (theor. monoisotopic ion at 203.1 m/z) derivative of the 13493.9 ± 0.2 m/z protein. On the other hand, the mass difference of 146 Da between the proteins with exp. monoisotopic [M+H]+ at 13843.1 ± 0.2 m/z and 13696.9 ± 0.2 m/z was in agreement with an additional deoxyhesose moiety (theor. monoisotopic ion at 146.1 m/z). Manual inspection of the high-resolution MS/MS spectra of the [M+10H]+10 ions at 1351.00, 1371.32 and 1386.11 m/z allowed to establish that the three proteins were different proteoforms of PIP with the N-terminal glutamine residue converted to pyro-glutamic acid and with two disulfide bonds, but not to assign the glycosylation site. The presence in the MS/MS spectra of low-molecular weight saccharide oxoniun ions at 204.087 and 138.106 m/z confirmed the presence of an N-acetylhexosamine moiety in the protein with [M+H]+ at 13696.9 ± 0.2 m/z.

2018-08-10 | PXD009440 | Pride

Salivary proteomics of sP and sNP rats

Project description:Sardinian alcohol-preferring (sP) and Sardinina alcohol-non preferring (sNP) rats have been selectively bred for opposite alcohol preference and consumption. The project proposed to identify salivary markers distinguishing the two rat lines possibly correlated to alcohol preference, by a proteomic approach.

2017-11-03 | PXD006997 | Pride

Human salivary basic proline-rich proteins

Project description:Human basic proline-rich proteins and basic glycosylated proline-rich proteins are the most complex family of human adult salivary proteins, secreted only by the parotid gland, expressed by the polymorphic PRB1-4 genes. This study reports the results obtained for their characterization in our laboratory utilizing integrated top-down and bottom-up HPLC-ESI-MS and MS/MS platform. The family include 11 parent peptides/proteins and more than 6 parent glycosylated proteins. Despite the complexity, on the basis of their primary structure they result the product of the duplication of three repeats and we propose that they can be classified in two main groups, the first including the peptides called P-E, P-Ko, IB-6, Ps-1, Ps-2, P-H, P-F, P-J, and P-D, the second including the peptides called IB-1, II-2 and the glycosylated basic proline-rich proteins codified by PRB3 and PRB4 genes. A third minor hybrid group includes the proteins called IB-8a Con1- and Con1+. Some of the proteins of the family (IB-1, II-2 and P-J) are almost always detectable in adult human saliva, while the others are expressed with different frequencies according to the phenotype. Various isoforms are detectable in the adult saliva and three of them, until now not reported (P-H S1→A, P-Ko P36→S, P-Ko A41→S), were characterized by MS/MS fragmentation.

2018-08-08 | PXD009813 | Pride

Human Salivary Cystatins - SALIVARY CYSTATINS: EXPLORING NEW POST-TRANSLATIONAL MODIFICATIONS AND POLYMORPHISMS BY TOP-DOWN HIGH RESOLUTION MASS SPECTROMETRY

Project description:In the present study various proteoforms of cystatin A, cystatin B, cystatin D, cystatin S, cystatin SN and cystatin SA were detected and characterized by a top-down HPLC-ESI-MS platform integrated by bottom-up strategies in the acidic soluble fraction of human saliva. The several proteoforms derive from polymorphisms of the coding sequence and post-translational modifications, typically phosphorylation, N-terminal processing, and oxidation. The protein species detected in saliva collected from healthy subjects were: (i) cystatin AT96→M and its acetylated derivative; (ii) cystatin B N-terminally acetylated and carboxymethylated at C3; (iii) N-terminally truncated cystatin D with the N-terminal Q converted to pyro-E and lacking the first 5 amino acid residues (pGlu-cystatin D Des1-5); (iv) N-terminally truncated forms of cystatin SN and SNP11→L lacking the first 4 amino acids (cystatin SN Des1-4 and cystatin SNP11→L Des1-4) and the first 7 amino acids (cystatin SN Des1-7 and cystatin SNP11→L Des1-7); (v) N-terminally truncated cystatin SA lacking the first 7 amino acids (cystatin SA Des1-7); (vi) oxidized derivatives of cystatins SN and S1 at W23 and W107.

2017-10-13 | PXD007170 | Pride

A transcriptomic resource for Douglas-fir seed development and analysis of transcription during late megagametophyte development

Project description:We developed a transcriptome resource for Douglas-fir covering key developmental stages of megagametophytes over time: prefertilization, fertilization, embryogenesis, and early, unfertilized abortion. Extracted RNA was sequenced using large-scale sequencing and reads were assembled to generate a de novo reference transcriptome of 105,505 predicted high-confidence transcripts. Expression levels were estimated based on alignment of the original reads to the reference. 200â400 megagametophytes were dissected and pooled per sample on four dates from either pollinated or unpollinated cones: June 10, June 22, June 30, and July 6 2011. These dates coincided with key events in seed development: corrosion cavity formation, fertilization, embryogenesis, or the early stages of abortion in the unpollinated treatment. Sporophytic tissue (i.e. cone bracts and cone scales) were added for comparison. PolyA RNA was used for Illumina sequencing.

2016-06-16 | E-GEOD-83425 | biostudies-arrayexpress

Site- and structure-specific glycosylation signatures of bovine, caprine, porcine and human milk-derived extracellular vesicles

Project description:Intact glycopeptides in MEVs from bovines, caprines, porcines and humans were comprehensively analyzed by high-resolution mass spectrometry using the sceHCD followed by EThcD fragment method, revealing that protein glycosylation is abundant and heterogeneous in MEVs.

2024-01-30 | PXD044641 | Pride

Deltamethrin resistance mechanisms in Aedes aegypti populations from three French overseas territories worldwide.

Project description:Custom microarrays were used to examine differential gene expression between pyrethroid resistant vs pyrethroid susceptible phenotypes of the dengue vector mosquito Aedes aegypti. Pyrethroid resistant population were from Cayenne (French Guiana, GUY), Baie Mahault (Guadeloupe, GUA) and Noumea (New Caledonia, CAL) whilst New Orleans lab colony represented the lab susceptible strain Pools of total RNA was extracted from the whole bodies of 3 day old female mosquitoes that had survived exposure to 0.06% deltamethrin (for GUY, GUA, CAL) . Single colour hybridization experiments were performed using labelled cDNA on the Agilent 'Aedes aegypti detox chip plus': A-MTAB-574. Four unique biological replicates per population were used in the study

2015-11-08 | E-MTAB-4022 | biostudies-arrayexpress

Tbeta4 identification CaCo2 cells

Project description:The paper describes new experiments about thymosin beta 4 nuclear translocation using different in vitro cellular stress in CaCo2 cell line adding new details regarding the role of this peptide during adaptive and defensive response. To this purpose, CaCo2 cells were submitted to different stress treatments, including serum starvation, DMSO and Butyrate treatment. The expression of Tβ4 was studied in culture cells before and after stress treatment in the cytosolic and nuclear fractions, with the use of different apporaches: immunohistochemistry with anti-Tβ4 antibody; mass-spectrometry characterization and quantification, and mRNA analysis.

2020-06-29 | PXD012161 | Pride

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