1. ¹Oxford Glycosciences (UK) Ltd, Abingdon, Oxford, UK
2. ²Academic Haematology, Institute of Cancer Research, Sutton, Surrey, UK
3. ³Department of Haematology, MRC Toxicology Unit/University of Leicester, UK
4. ⁴Department of Haematology, Royal Bournemouth Hospital, UK

Correspondence: Professor Dr MJS Dyer, Department of Haematology, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary PO Box 65, Leicester, LE2 7LX, UK. Fax: +44 116 252 3274

⁵These authors contributed equally to the work presented in this paper

Abstract

B-cell-specific plasma-membrane proteins are potential targets for either small molecule or antibody-based therapies. We have sought to annotate proteins expressed at the cell surface membrane in patients with chronic lymphocytic leukemia (CLL) using plasma-membrane-based proteomic analysis to identify previously uncharacterized and potentially B-cell-specific proteins. Proteins from plasma-membrane fractions were separated on one-dimensional gels and trypsinized fractions subjected to high-throughput MALDI-TOF mass spectrometry. Using this method, many known B-cell surface antigens were detected, but also known proteins not previously described in this disease or in this cellular compartment, including cell surface receptors, membrane-associated enzymes and secreted proteins, and completely unknown proteins. To validate the method, we show that BLK, a B-cell-specific kinase, is located in the CLL-plasma-membrane fraction. We also describe two novel proteins (MIG2B and B-cell novel protein #1, BCNP1), which are expressed preferentially in B cells. MIG2B is in a highly conserved and defined gene family containing two plasma-membrane-binding ezrin/radixin/moesin domains and a pleckstrin homology domain; the Caenorhabditis elegans homolog (UNC-112) is a membrane-associated protein that colocalizes with integrin at cell–matrix adhesion complexes. BCNP1 is a completely unknown protein with three predicted transmembrane domains, with three alternatively spliced final exons. Proteomic analysis may thus define new potential therapeutic targets.