Dataset Information

A label-free mass spectrometry method for relative quantitation of ?-tubulin isotype expression in human tumor tissue.

ABSTRACT: Quantitation of ?-tubulin isotype expression in taxane resistant human tumor tissue has been difficult to achieve because of the limited availability of validated antibodies. Here we present a label-free MS method to quantitate relative expression levels of ?-tubulin isotypes.Using isotype-specific reporter peptides, we determined relative ?-tubulin isotype expression levels in human lung tumor tissue.Four reporter peptides were chosen to quantitate the ?I/?II, ?IV, ?III, and ?V tubulin isotypes. These peptides were validated using human cancer cell lines. The label-free method was then used to determine ?-tubulin isotype expression in nine human lung tumor samples, which had been described as high or low ?III-tubulin expressing using immunohistochemistry. It was found that ?I/?II (accounting for 18.7-65.7% of total ?-tubulin) and ?IVa/?IVb (26.3-79.1%) were the most abundant isotypes and that the ?III (0-8.9%) and ?V (1.0-10.4%) were less abundant in the tissue. We also categorized the samples as high or low ?III-tubulin expressing.With this method we can determine the relative expression levels of ?-tubulin isotypes in human tumor tissue. This method will facilitate studies assessing the use of tubulin isotypes as biomarkers of taxane resistance.

SUBMITTER: Miller LM

PROVIDER: S-EPMC3484181 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue.

Miller Leah M LM Huang Yang Chia-Ping CP Xiao Hui H Isaac Sylvie S Sève Pascal P Dumontet Charles C Band Horwitz Susan S Hogue Angeletti Ruth R

Proteomics. Clinical applications 20121001 9-10

<h4>Purpose</h4>Quantitation of β-tubulin isotype expression in taxane resistant human tumor tissue has been difficult to achieve because of the limited availability of validated antibodies. Here we present a label-free MS method to quantitate relative expression levels of β-tubulin isotypes.<h4>Experimental design</h4>Using isotype-specific reporter peptides, we determined relative β-tubulin isotype expression levels in human lung tumor tissue.<h4>Results</h4>Four reporter peptides were chosen ...[more]

PMID: 22996942

Similar Datasets

Project description:BACKGROUND: Label-free quantitation of mass spectrometric data is one of the simplest and least expensive methods for differential expression profiling of proteins and metabolites. The need for high accuracy and performance computational label-free quantitation methods is still high in the biomarker and drug discovery research field. However, recent most advanced types of LC-MS generate huge amounts of analytical data with high scan speed, high accuracy and resolution, which is often impossible to interpret manually. Moreover, there are still issues to be improved for recent label-free methods, such as how to reduce false positive/negatives of the candidate peaks, how to expand scalability and how to enhance and automate data processing. AB3D (A simple label-free quantitation algorithm for Biomarker Discovery in Diagnostics and Drug discovery using LC-MS) has addressed these issues and has the capability to perform label-free quantitation using MS1 for proteomics study. RESULTS: We developed an algorithm called AB3D, a label free peak detection and quantitative algorithm using MS1 spectral data. To test our algorithm, practical applications of AB3D for LC-MS data sets were evaluated using 3 datasets. Comparisons were then carried out between widely used software tools such as MZmine 2, MSight, SuperHirn, OpenMS and our algorithm AB3D, using the same LC-MS datasets. All quantitative results were confirmed manually, and we found that AB3D could properly identify and quantify known peptides with fewer false positives and false negatives compared to four other existing software tools using either the standard peptide mixture or the real complex biological samples of Bartonella quintana (strain JK31). Moreover, AB3D showed the best reliability by comparing the variability between two technical replicates using a complex peptide mixture of HeLa and BSA samples. For performance, the AB3D algorithm is about 1.2 - 15 times faster than the four other existing software tools. CONCLUSIONS: AB3D is a simple and fast algorithm for label-free quantitation using MS1 mass spectrometry data for large scale LC-MS data analysis with higher true positive and reasonable false positive rates. Furthermore, AB3D demonstrated the best reproducibility and is about 1.2- 15 times faster than those of existing 4 software tools.

Dataset Information

A label-free mass spectrometry method for relative quantitation of ?-tubulin isotype expression in human tumor tissue.

Publications

A label-free mass spectrometry method for relative quantitation of β-tubulin isotype expression in human tumor tissue.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets