From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia |
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Authors: | Benedetta Apollonio Maria Teresa Sabrina Bertilaccio Umberto Restuccia Pamela Ranghetti Federica Barbaglio Paolo Ghia Federico Caligaris-Cappio Cristina Scielzo |
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Institution: | 1Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute;2Department of Haemato-Oncology, King''s College London;3IFOM, FIRC Institute of Molecular Oncology;4Università Vita-Salute San Raffaele |
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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. |
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Keywords: | Medicine Issue 92 Lymphocytes Chronic Lymphocytic Leukemia 2D Electrophoresis Mass Spectrometry Cytoskeleton Migration |
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