Identification of the key LMO2-binding determinants on Ldb1

Journal of Molecular Biology
Daniel P RyanJacqueline M Matthews

Abstract

The overexpression of LIM-only protein 2 (LMO2) in T-cells, as a result of chromosomal translocations, retroviral insertion during gene therapy, or in transgenic mice models, leads to the onset of T-cell leukemias. LMO2 comprises two protein-binding LIM domains that allow LMO2 to interact with multiple protein partners, including LIM domain-binding protein 1 (Ldb1, also known as CLIM2 and NLI), an essential cofactor for LMO proteins. Sequestration of Ldb1 by LMO2 in T-cells may prevent it binding other key partners, such as LMO4. Here, we show using protein engineering and enzyme-linked immunosorbent assay (ELISA) methodologies that LMO2 binds Ldb1 with a twofold lower affinity than does LMO4. Thus, excess LMO2 rather than an intrinsically higher binding affinity would lead to sequestration of Ldb1. Both LIM domains of LMO2 are required for high-affinity binding to Ldb1 (K(D) = 2.0 x 10(-8) M). However, the first LIM domain of LMO2 is primarily responsible for binding to Ldb1 (K(D) = 2.3 x 10(-7) M), whereas the second LIM domain increases binding by an order of magnitude. We used mutagenesis in combination with yeast two-hybrid analysis, and phage display selection to identify LMO2-binding "hot spots" within Ldb1 that locate t...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·T A KunkelJ McClary
Oct 10, 1995·Proceedings of the National Academy of Sciences of the United States of America·H OsadaT H Rabbitts
Jan 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·S Jones, J M Thornton
Feb 1, 1996·Journal of Molecular Graphics·R KoradiK Wüthrich
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·L W JurataG N Gill
Oct 7, 1997·Molecular and Cellular Biology·L W Jurata, G N Gill
Feb 12, 1998·Proceedings of the National Academy of Sciences of the United States of America·J E VisvaderS H Orkin
May 9, 1998·Proceedings of the National Academy of Sciences of the United States of America·Y YamadaT H Rabbitts
Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·C ZengY N Jan
Jan 5, 2000·Proceedings of the National Academy of Sciences of the United States of America·Y YamadaT H Rabbitts
Mar 8, 2000·Mechanisms of Development·I Bach
Nov 15, 2000·Methods in Enzymology·S S SidhuJ A Wells
Jun 18, 2002·FEBS Letters·Valérie Bégay-MüllerAchim Leutz
May 3, 2003·The EMBO Journal·Janet E DeaneJacqueline M Matthews
Jun 26, 2003·Nucleic Acids Research·Torsten SchwedeManuel C Peitsch
Dec 4, 2003·EMBO Reports·Jacqueline M Matthews, Jane E Visvader
Sep 3, 2004·The EMBO Journal·Janet E DeaneJacqueline M Matthews
Apr 5, 2005·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Eleanor Y M SumJane E Visvader
Nov 2, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chang-Hoon Nam, Terence H Rabbitts

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Citations

Jun 22, 2010·Biomolecular NMR Assignments·Lorna E Wilkinson-WhiteJacqueline M Matthews
Jun 7, 2011·Biomolecular NMR Assignments·Chu Wai LiewJacqueline M Matthews
Jan 11, 2013·Nature Reviews. Cancer·Jacqueline M MatthewsDavid J Curtis
Aug 17, 2011·Proceedings of the National Academy of Sciences of the United States of America·Lorna Wilkinson-WhiteJacqueline M Matthews
Apr 12, 2012·Nucleic Acids Research·K BendakJ P Mackay
Nov 4, 2010·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Kamel El OmariErika J Mancini
Oct 3, 2007·Proteins·Daniel P RyanJacqueline M Matthews
Sep 1, 2012·Protein Science : a Publication of the Protein Society·Siavoush DastmalchiJacqueline M Matthews
Jun 26, 2015·Open Biology·Jennifer Chambers, Terence H Rabbitts
Jun 10, 2008·Developmental Cell·Seunghee LeeSoo-Kyung Lee
Sep 21, 2016·Angewandte Chemie·Neil O RobertsonJacqueline M Matthews
Nov 26, 2015·Molecular and Cellular Biology·Justin H LayerUtpal P Davé
Jan 22, 2013·FEBS Letters·Jacqueline M Matthews, Jennifer R Potts
Jan 18, 2008·Journal of Molecular Recognition : JMR·Oksana Okhrimenko, Ilian Jelesarov
Dec 13, 2007·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Nov 22, 2008·Biochemical Society Transactions·Jacqueline M MatthewsMargaret Sunde
Jun 12, 2013·The Journal of Biological Chemistry·Morgan S GaddJacqueline M Matthews

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