The structure-function relationship of oncogenic LMTK3
Authors: Storm Hassell-Hart
Raymond J Owens
Georgios Giamas
S Mark Roe
Apostolos Klinakis
Lihong Zhou
Chiara Cilibrasi
Goar Smbatyan
Thomas Simon
Leanne Milton-Harris
Shivani Soni
Angeliki Ditsiou
Jae Ho Lo
Maria Chiara Iachini
John Spencer
Viviana Vella
Heinz-Josef Lenz
Naomi E Chayen
Teresa Gagliano
Panagiota Ntavelou
Chrisostomos Prodromou
Robin L Owen
Justin Stebbing
Sahir Khurshid
Laurence H Pearl
Nikiana Simigdala
Joanne E Nettleship
Philip Carter
Athanasios Papakyriakou
Journal: Science Advances (2020)
Discipline: Multidisciplinary
Keywords: LMTK3; kinase domain; crystal structure; phosphoproteome; small-molecule inhibitor
Affiliations: Department of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK., Center of Basic Research, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece., Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos," 15341 Athens, Greece., Division of Structural Biology, University of Oxford, The Wellcome Centre for Human Genetics Headington, Oxford OX3 7BN, UK., Protein Production UK, Research Complex at Harwell, Harwell Science and Innovation Campus, Didcot OX11 0FA, UK., Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA., Faculty of Medicine, Division of Systems Medicine, Department of Metabolism, Digestion and Reproduction, Imperial College, Sir Alexander Fleming Building, South Kensington Campus, London SW7 2AZ, UK., Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Science Park Road, Falmer, Brighton BN1 9RQ, UK., Department of Chemistry, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QJ, UK., Faculty of Medicine, Department of Surgery and Cancer, Imperial College, London W12 0NN, UK., Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK., The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0FA, UK., Department of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK. .

Abstract:  Elucidating signaling driven by lemur tyrosine kinase 3 (LMTK3) could help drug development. Here, we solve the crystal structure of LMTK3 kinase domain to 2.1√Ö resolution, determine its consensus motif and phosphoproteome, unveiling in vitro and in vivo LMTK3 substrates. Via high-throughput homogeneous time-resolved fluorescence screen coupled with biochemical, cellular, and biophysical assays, we identify a potent LMTK3 small-molecule inhibitor (C28). Functional and mechanistic studies reveal LMTK3 is a heat shock protein 90 (HSP90) client protein, requiring HSP90 for folding and stability, while C28 promotes proteasome-mediated degradation of LMTK3. Pharmacologic inhibition of LMTK3 decreases proliferation of cancer cell lines in the NCI-60 panel, with a concomitant increase in apoptosis in breast cancer cells, recapitulating effects of LMTK3 gene silencing. Furthermore, LMTK3 inhibition reduces growth of xenograft and transgenic breast cancer mouse models without displaying systemic toxicity at effective doses. Our data reinforce LMTK3 as a druggable target for cancer therapy. 

Retracted: No

PubPeer comments:
 The Tubulin bands used as loading controls for T47D cells and MDA-MB-231 cells in Figure 5D look more similar than expected (red boxes below). Could the authors comment?  , 
 This paper appears to have undergone a stealth correction, with the replacement of the offending repeated western panel in Figure 5D.  , 
 Near the supplemental information: u201cCorrection (2 April 2021): In Fig. 5D, the tubulin bands for T47D and MDA-MB-231 cells were inadvertently identical. The PDF and HTML have been updated with the correct version of Figure 5.u201d , 
 Fortunately, for history s sake, the  original version continues to exist on PubMed: https: www.ncbi.nlm.nih.gov pmc articles PMC7673765 pdf abc3099.pdf ,