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Validation of an LC‐MS based approach for profiling histones in chronic lymphocytic leukemia |
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| Authors: | Xiaodan Su David M Lucas Liwen Zhang Hua Xu Vlad Zabrouskov Melanie E Davis Amy R Knapp Donn C Young Philip R O Payne Mark R Parthun Guido Marcucci Michael R Grever John C Byrd Michael A Freitas Dr |
| Institution: | 1. Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH, USA;2. Hematology and Oncology, The Ohio State University, Columbus, OH, USA;3. Campus Chemical Instrument Center, The Ohio State University, Columbus, OH, USA;4. CBC/RRC Proteomics & Informatics Services Facility, University of Illinois at Chicago, Chicaga, IL, USA;5. Thermo Fisher Scientific, San Jose, CA, USA;6. Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH, USA;7. Center for Biostatistics, The Ohio State University, Columbus, OH, USA;8. Biomedical Informatics, The Ohio State University, Columbus, OH, USA |
| Abstract: | The in vitro evaluation of histones and their PTMs has drawn substantial interest in the development of epigenetic therapies. The differential expression of histone isoforms may serve as a potential marker in the classification of diseases affected by chromatin abnormalities. In this study, protein profiling by LC and MS was used to explore differences in histone composition in primary chronic lymphocytic leukemia (CLL) cells. Extensive method validations were performed to determine the experimental variances that would impact histone relative abundance. The resulting data demonstrated that the proposed methodology was suitable for the analysis of histone profiles. In 4 normal individuals and 40 CLL patients, a significant decrease in the relative abundance of histone H2A variants (H2AFL and H2AFA/M*) was observed in primary CLL cells as compared to normal B cells. Protein identities were determined using high mass accuracy MS and shotgun proteomics. |
| Keywords: | Chronic lymphocytic leukemia Histone RPLC‐MS Variant |