1. From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia
- Author
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Umberto Restuccia, Benedetta Apollonio, Federica Barbaglio, Cristina Scielzo, Federico Caligaris-Cappio, Paolo Ghia, Pamela Ranghetti, Maria Teresa Sabrina Bertilaccio, Apollonio, B, Bertilaccio, Mt, Restuccia, U, Ranghetti, P, Barbaglio, F, Ghia, PAOLO PROSPERO, Caligaris Cappio, F, and Scielzo, C.
- Subjects
Proteomics ,Chronic lymphocytic leukemia ,General Chemical Engineering ,Molecular Sequence Data ,Tumor initiation ,General Biochemistry, Genetics and Molecular Biology ,Cell Movement ,medicine ,Humans ,Electrophoresis, Gel, Two-Dimensional ,Amino Acid Sequence ,Phosphorylation ,Cytoskeleton ,Peptide sequence ,Adaptor Proteins, Signal Transducing ,Protein function ,B-Lymphocytes ,Two-dimensional gel electrophoresis ,General Immunology and Microbiology ,business.industry ,General Neuroscience ,Blood Proteins ,medicine.disease ,Molecular biology ,Leukemia, Lymphocytic, Chronic, B-Cell ,Actins ,Leukemia ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Immunology ,Medicine ,business - Abstract
The identification of molecules involved in tumor initiation and progression is fundamental for understanding disease’s biology and, as a consequence, for the clinical management of patients. In the present work we will describe an optimized proteomic approach for the identification of molecules involved in the progression of Chronic Lymphocytic Leukemia (CLL). In detail, leukemic cell lysates are resolved by 2-dimensional Electrophoresis (2DE) and visualized as “spots” on the 2DE gels. Comparative analysis of proteomic maps allows the identification of differentially expressed proteins (in terms of abundance and post-translational modifications) that are picked, isolated and identified by Mass Spectrometry (MS). The biological function of the identified candidates can be tested by different assays (i.e. migration, adhesion and F-actin polymerization), that we have optimized for primary leukemic cells.
- Published
- 2014
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