Dataset Information

Targeted Quantitative Proteomic Approach for High-Throughput Quantitative Profiling of Small GTPases in Brain Tissues of Alzheimer's Disease Patients.

ABSTRACT: Neurodegenerative disorders, including Alzheimer's disease (AD), are prevalent among the elderly. Small GTPases of the Ras superfamily are essential regulators of intracellular trafficking and signal transduction. In this study, we develop a targeted quantification method for small GTPase proteins, where the method involves scheduled multiple-reaction monitoring analysis and the use of synthetic stable isotope-labeled peptides as internal standards or surrogate standards. We further applied this method to examine the altered expression of small GTPase proteins in post-mortem frontal cortex tissues from AD patients with different degrees of disease severity. We were able to achieve sensitive and reproducible quantifications of 80 small GTPases in brain tissue samples from 15 patients. Our results revealed substantial up-regulations of several synaptic GTPases, i.e., RAB3A/C, RAB4A/B, and RAB27B, in tissues from patients with higher degrees of AD pathology, suggesting that aberrant synaptic trafficking may modulate the progression of AD. The method should be generally applicable for high-throughput targeted quantification of small GTPase proteins in other tissue and cellular samples.

SUBMITTER: Huang M

PROVIDER: S-EPMC6939614 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Targeted Quantitative Proteomic Approach for High-Throughput Quantitative Profiling of Small GTPases in Brain Tissues of Alzheimer's Disease Patients.

Huang Ming M Darvas Martin M Keene C Dirk CD Wang Yinsheng Y

Analytical chemistry 20190910 19

Neurodegenerative disorders, including Alzheimer's disease (AD), are prevalent among the elderly. Small GTPases of the Ras superfamily are essential regulators of intracellular trafficking and signal transduction. In this study, we develop a targeted quantification method for small GTPase proteins, where the method involves scheduled multiple-reaction monitoring analysis and the use of synthetic stable isotope-labeled peptides as internal standards or surrogate standards. We further applied this ...[more]

PMID: 31460748

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Project description:BackgroundThe recent development within high-throughput technologies for expression profiling has allowed for parallel analysis of transcriptomes and proteomes in biological systems such as comparative analysis of transcript and protein levels of tissue regulated genes. Until now, such studies of have only included microarray or short length sequence tags for transcript profiling. Furthermore, most comparisons of transcript and protein levels have been based on absolute expression values from within the same tissue and not relative expression values based on tissue ratios.ResultsPresented here is a novel study of two porcine tissues based on integrative analysis of data from expression profiling of identical samples using cDNA microarray, 454-sequencing and iTRAQ-based proteomics. Sequence homology identified 2.541 unique transcripts that are detectable by both microarray hybridizations and 454-sequencing of 1.2 million cDNA tags. Both transcript-based technologies showed high reproducibility between sample replicates of the same tissue, but the correlation across these two technologies was modest. Thousands of genes being differentially expressed were identified with microarray. Out of the 306 differentially expressed genes, identified by 454-sequencing, 198 (65%) were also found by microarray. The relationship between the regulation of transcript and protein levels was analyzed by integrating iTRAQ-based proteomics data. Protein expression ratios were determined for 354 genes, of which 148 could be mapped to both microarray and 454-sequencing data. A comparison of the expression ratios from the three technologies revealed that differences in transcript and protein levels across heart and muscle tissues are positively correlated.ConclusionWe show that the reproducibility within cDNA microarray and 454-sequencing is high, but that the agreement across these two technologies is modest. We demonstrate that the regulation of transcript and protein levels across identical tissue samples is positively correlated when the tissue expression ratios are used for comparison. The results presented are of interest in systems biology research in terms of integration and analysis of high-throughput expression data from mammalian tissues.

Dataset Information

Targeted Quantitative Proteomic Approach for High-Throughput Quantitative Profiling of Small GTPases in Brain Tissues of Alzheimer's Disease Patients.

Publications

Targeted Quantitative Proteomic Approach for High-Throughput Quantitative Profiling of Small GTPases in Brain Tissues of Alzheimer's Disease Patients.

Similar Datasets

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