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Human testis off-line LC-MS/MS

ABSTRACT: In order to identify more MPs and get deeper proteomic data of testis. We applied high-pH reverse phase (RP) HPLC to fractionate complex samples.On the other hand, to enhance protein coverage, multi-protease strategies were used for 10% SDS-PAGE short separation samples.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Ping Xu

PROVIDER: MSV000086491 | MassIVE | Thu Nov 19 23:03:00 GMT 2020

SECONDARY ACCESSION(S): PXD009737

REPOSITORIES: MassIVE

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Multiproteases Combined with High-pH Reverse-Phase Separation Strategy Verified Fourteen Missing Proteins in Human Testis Tissue.

Sun Jinshuai J Shi Jiahui J Wang Yihao Y Chen Yang Y Li Yanchang Y Kong Degang D Chang Lei L Liu Fengsong F Lv Zhitang Z Zhou Yue Y He Fuchu F Zhang Yao Y Xu Ping P

Journal of proteome research 20181016 12

Subsequent to conducting the Chromosome-Centric Human Proteome Project, we have focused on human testis-enriched missing proteins (MPs) since 2015. For protein coverage to be enhanced, a multiprotease strategy was used for separation of samples by 10% SDS-PAGE. For the separating efficiency to be improved, a high-pH reverse phase (RP) separation strategy was applied to fractionate complex samples in this study. A total of 11,558 proteins was identified, which is the largest proteome data set for ...[more]

PMID: 30280576

Similar Datasets

Human testis off-line LC-MS/MS

Project description:In order to identify more MPs and get deeper proteomic data of testis. We applied high-pH reverse phase (RP) HPLC to fractionate complex samples.On the other hand, to enhance protein coverage, multi-protease strategies were used for 10% SDS-PAGE short separation samples.

2018-10-04 | PXD009737 | Pride

CN-CHPP 2015 Testis Project

Project description:The testis tissue-based trans-omics was a feasible approach to find more missing proteins (MPs) because it was reported higher gene expression found in testis tissues. Three testis tissues and two protein separation strategies were selected to quantitatively evaluate gene expression abundance at the proteomic and transcriptomic levels. The project have been jointly supervised by Ping Xu and Siqi Liu.

2018-06-26 | MSV000082510 | MassIVE

CN-CHPP 2015 Testis Project - Tissue-Based Proteogenomics Reveals that Human Testis Endows Plentiful Missing Proteins

2015-11-03 | PXD002179 | Pride

Human Testis LC-MS/MS

Project description:Missing proteins(MPs) investigation is one of the critical missions of Chromosome-based Human Proteome Project(C-HPP). Through analyzing sequence of MPs from neXtProt database, we found that there are no detectable tryptic peptides for some MPs. Here, we attmept a multi-enzymes strategy to cover this problem. Lys-C, Glu-C and LysargiNase are used to produce more unique and detectable peptides of MPs.

2019-06-17 | MSV000083982 | MassIVE

Human Testis LC-MS/MS - Multi-Protease Strategy Identifies Three PE2 Missing Proteins in Human Testis Tissue

2017-10-09 | PXD006465 | Pride

Human_Testis_QE-HF - Deep Coverage Proteomics Identifies More Low-Abundance Missing Proteins in Human Testis Tissue with Q-Exactive HF Mass Spectrometer

Project description:The Chromosome-Centric Human Proteome Project (C-HPP) aims at finding existence evidence of missing proteins (MPs) at the early stage. Based on previous research, it has been proved that human testis tissue is suitable for further research of MPs because of its high gene expression level and high transcriptome complexity. However, the limits of dynamic range in mass spectrometric analysis and the deficiency in sensitivity and resolution will seriously affect the detection efficiency. In this study, we used Q-Exactive HF to detect three individual testis tissues dealted with two separation methods (tricine-SDS-PAGE and SDS-PAGE).

2016-09-30 | PXD004092 | Pride

Silkworm testis LC-MS/MS - Proteomics analysis of adult testis from Bombyx mori

Project description:The development of the testis involves a large number of tissue-specific proteins, possibly because that sperms in it are the most divergent of all cell types. Although the proteome of the larval testis of Bombyx mori has been reported, no comprehensive data on adult testis proteome is available for silkworm. In this study, Liquid chromatography-tandem mass spectrometry was employ to investigate the protein compositions of the adult testis of silkworm. A total of 14431 peptides were identified in the adult testis of B. mori, which were match to 2292 proteins. Thirty-two heat shock proteins constitute a group of most abundant proteins in the adult testis, suggested that they are critical for the development, differentiation and survival of germ cells. Other proteins in this analysis were also involved in testis-specific processes mainly including sperm motility, meiosis, germ cell development, and spermatogenesis.

2014-07-28 | PXD000909 | Pride

Buffalo testis ubiquitome LC-MS/MS

Project description:Protein ubiquitination, a major and conserved post-translational modification, is known to play a critical regulatory role in many biological processes in eukaryotes. Although several ubiquitinated proteins have been found in buffalo (Bubalus bubalis) testis in our previous studies, large-scale profiling of buffalo testis ubiquitome has not been reported to date. In this study, we firstly identified a global profiling of lysine ubiquitination of adult buffalo testis using a highly sensitive LC-MS/MS coupled with immune-affinity enrichment of ubiquitinated peptides. In total, 422 lysine ubiquitination sites were identified in 262 proteins in adult buffalo testis tissue. Bioinformatic analysis showed that the ubiquitinated proteins are involved in a variety of biological processes and diverse subcellular localizations. Besides, KEGG pathway and protein interaction network analysis indicated that proteasome, glycolysis/gluconeogenesis and gap junction pathways are modulated by protein ubiquitination in testis. Taken together, these data provide a global view of ubiquitome in buffalo testis for the first time, and serve as an important resource for exploring the physiological role of lysine ubiquitination in testis in mammalian.

2020-06-05 | PXD012231 | Pride

Molecular Elucidation of in vitro spermatogenesis in the testis organ culture

Project description:Cultivation of neonatal mouse testis tissue can induce spermatogenesis and produce fertile sperms. However, this in vitro spermatogenesis by the current organ culture method comes short in fully mimicking the in vivo counterpart, partly due to a lack of knowledge about the underlying molecular phenotypes. In this study, we investigated transcriptome of cultured testis tissues using microarray method. Principle component analysis of the transcriptome data revealed delay and/or arrest of spermatogenesis and immediate radical immune reactions in the cultured testis tissues. The delay/arrest of spermatogenesis occurred before and during early meiotic phase, resulting in inefficient progression of meiosis. The immune reaction, on the other hand, was drastic and overwhelming, in which TLR4-NF-kB signaling was speculated to be involved. Notably, treatment with TAK242, an inhibitor of TLR4-NF-kB signaling pathway, ameliorated the macrophage activation which otherwise would exacerbate the inflammation. The present study revealed for the first time the immense immune reaction in the cultured tissue by transcriptomic analysis, which could be a cause of the inefficiency of in vitro spermatogenesis.

2020-06-30 | GSE147982 | GEO

A shot to missing protein from an aneuploidy cell line

Project description:With fast development of proteomic technology, the scale of missing proteins (MPs) has being continuously shrunk, approximately 1,470 MPs not explored yet. Discovery of MPs, on the other hand, is appearing more difficult. In some aneuploid cells, the abundance changes are proportional to the additional chromosome(s), while several MPs were found within them. Herein, we hypothesize that a stable aneuploid cell line with the increased chromosomes was a useful material that assists MP exploration. Ker-CT cell line with trisomy at chromosome 5 and 20 was selected. With a combination strategy of RNA-Seq and LC-M/MS, a total of 22,178 transcripts and 8,846 proteins were identified in Ker-CT. Although the transcripts corresponding to 31 and 32 MP genes located at chromosome 5 and 20 were detected, none of MPs was found in Ker-CT. Surprisingly, 3 MPs containing at least two unique non-nest peptides of length ≥9 amino acid were identified in Ker-CT, whose genes are located in chromosome 3 and 10. Furthermore, the 3 MPs were verified using the method of parallel reaction monitoring (PRM).

2021-11-25 | PXD028647 | Pride

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