title: From a 2DE-gel spot to protein function: lesson learned from HS1 in chronic lymphocytic leukemia.
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.
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute; Department of Haemato-Oncology, King's College London.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Apollonio
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute.
authors.forename: Maria Teresa Sabrina
authors.initials: MT
authors.lastname: Bertilaccio
authors.affiliation: IFOM, FIRC Institute of Molecular Oncology.
authors.forename: Umberto
authors.initials: U
authors.lastname: Restuccia
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute.
authors.forename: Pamela
authors.initials: P
authors.lastname: Ranghetti
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute.
authors.forename: Federica
authors.initials: F
authors.lastname: Barbaglio
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute; Università Vita-Salute San Raffaele.
authors.forename: Paolo
authors.initials: P
authors.lastname: Ghia
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute; Università Vita-Salute San Raffaele.
authors.forename: Federico
authors.initials: F
authors.lastname: Caligaris-Cappio
authors.affiliation: Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute; scielzo.cristina@hsr.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Scielzo
mesh_terms: D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000473:pathology; D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology; D003599:Cytoskeleton / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000097:blood / Q000378:metabolism* / Q000473:pathology; D008969:Molecular Sequence Data; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins; C061909:HCLS1 protein, human