Structural basis of p38α regulation by hematopoietic tyrosine phosphatase

Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode Island, USA.
Nature Chemical Biology (Impact Factor: 13). 11/2011; 7(12):916-924. DOI: 10.1038/nchembio.707
Source: PubMed


MAP kinases regulate essential cellular events, including cell growth, differentiation and inflammation. The solution structure of a complete MAPK-MAPK-regulatory protein complex, p38α-HePTP, was determined, enabling a comprehensive investigation of the molecular basis of specificity and fidelity in MAPK regulation. Structure determination was achieved by combining NMR spectroscopy and small-angle X-ray scattering data with a new ensemble calculation-refinement procedure. We identified 25 residues outside of the HePTP kinase interaction motif necessary for p38α recognition. The complex adopts an extended conformation in solution and rarely samples the conformation necessary for kinase deactivation. Complex formation also does not affect the N-terminal lobe, the activation loop of p38α or the catalytic domain of HePTP. Together, these results show how the downstream tyrosine phosphatase HePTP regulates p38α and provide for fundamentally new insights into MAPK regulation and specificity.

Download full-text


Available from: Bartosz Różycki,
  • Source
    • "This is of special interest, as recent studies have shown that while KIM-peptides derived from KIM-containing regulatory proteins and substrates readily discriminate between ERK2/p38 and JNK, they discriminate poorly between ERK2 and p38 [7]. We recently reported detailed ITC, NMR as well as small angle X-ray scattering (SAXS) data of the interactions between the MAPK p38α and the KIM-PTP family members [14], [15]. Furthermore, we also compared the interaction the immune-specific KIM-PTP, HePTP, with p38α and ERK2 [16], [17]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The mitogen-activation protein kinase ERK2 is tightly regulated by multiple phosphatases, including those of the kinase interaction motif (KIM) PTP family (STEP, PTPSL and HePTP). Here, we use small angle X-ray scattering (SAXS) and isothermal titration calorimetry (ITC) to show that the ERK2:STEP complex is compact and that residues outside the canonical KIM motif of STEP contribute to ERK2 binding. Furthermore, we analyzed the interaction of PTPSL with ERK2 showing that residues outside of the canonical KIM motif also contribute to ERK2 binding. The integration of this work with previous studies provides a quantitative and structural map of how the members of a single family of regulators, the KIM-PTPs, differentially interact with their corresponding MAPKs, ERK2 and p38α.
    PLoS ONE 03/2014; 9(3):e91934. DOI:10.1371/journal.pone.0091934 · 3.23 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: This chapter highlights a selection of the most recent publications applying NMR spectroscopy to the field of structural biology. The reports are split approximately into the general areas of methods development (section 2, "New methodology") and applications (sections 3 and 4, "High-resolution structural studies" and "NMR studies of large biomolecules", respectively). As the majority of new developments are associated with either sample production and/or improved software, this year all the new methodology reports have been incorporated into two general sections; (2.1) "Samples" and 2.2 "Software, web tools and protocols". The unparalleled level and variety of data that NMR can provide on biological systems means that there is an ever increasing number of high-quality studies reported annually. Hence, this review only provides a selection of particularly interesting publications or challenging studies, with the latter part devoted to large biomolecules (section 4, split arbitrarily into "Large, soluble molecules"; 4.1, "Membrane proteins"; 4.2 and "Very large systems"; 4.3).
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The MAP kinase ERK2 (ERK2, extracellular signal-regulated kinase 2) is regulated by numerous phosphatases that tightly control its activity. For example, the hematopoietic tyrosine phosphatase (HePTP) negatively regulates T cell activation in lymphocytes via ERK2 dephosphorylation. However, only very limited structural information is available for these biologically important complexes. Here, we use small-angle X-ray scattering combined with EROS ensemble refinement to characterize the structures of the resting and active states of ERK2:HePTP complexes. Our data show that the resting state ERK2:HePTP complex adopts a highly extended, dynamic conformation that becomes compact and ordered in the active state complex. This work experimentally demonstrates that these complexes undergo significant dynamic structural changes in solution and provides the first structural insight into an active state MAPK complex.
    Journal of the American Chemical Society 11/2011; 133(43):17138-41. DOI:10.1021/ja2075136 · 12.11 Impact Factor
Show more