The Crystal Structure of a Constitutively Active Mutant RON Kinase Suggests an Intramolecular Autophosphorylation Hypothesis

Wang, Jing; Steinbacher, Stefan; Augustin, Martin; Schreiner, Patrick; Epstein, David; Mulvihill, Mark J.; Crew, Andrew P.

doi:10.1021/bi100409w

Cited by 18 publications

(21 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7D). We also superposed structures of RON [PDB: 3PLS (79)] and MET [PDB: 1R0P (80)], which contain Tyr 1353 and Tyr 1349 residues, respectively, at homologous positions to Tyr 766 of FGFR1. The superposition showed that in the structures in which the Tyr is not bound to another protein (either kinase or PLCγ), the C-terminal Tyr residues are in the same position up against the E helix.…”

Section: Resultsmentioning

confidence: 99%

Identifying three-dimensional structures of autophosphorylation complexes in crystals of protein kinases

Malecka

Fink

et al. 2015

Sci. Signal.

View full text Add to dashboard Cite

Protein kinase autophosphorylation is a common regulatory mechanism in cell signaling pathways. Crystal structures of several homomeric protein kinase complexes have a serine, threonine, or tyrosine autophosphorylation site of one kinase monomer located in the active site of another monomer, a structural complex that we call an “autophosphorylation complex.” We developed and applied a structural bioinformatics method to identify all such autophosphorylation kinase complexes in X-ray crystallographic structures in the Protein Data Bank (PDB). We identified 15 autophosphorylation complexes in the PDB, of which 5 complexes had not previously been described in the publications describing the crystal structures. These 5 consist of tyrosine residues in the N-terminal juxtamembrane regions of colony stimulating factor 1 receptor (CSF1R, Tyr561) and EPH receptor A2 (EPHA2, Tyr594), tyrosine residues in the activation loops of the SRC kinase family member LCK (Tyr394) and insulin-like growth factor 1 receptor (IGF1R, Tyr1166), and a serine in a nuclear localization signal region of CDC-like kinase 2 (CLK2, Ser142). Mutations in the complex interface may alter autophosphorylation activity and contribute to disease; therefore we mutated residues in the autophosphorylation complex interface of LCK and found that two mutations impaired autophosphorylation (T445V and N446A) and mutation of Pro447 to Ala, Gly, or Leu increased autophosphorylation. The identified autophosphorylation sites are conserved in many kinases, suggesting that, by homology, these complexes may provide insight into autophosphorylation complex interfaces of kinases that are relevant drug targets.

show abstract

Section: Resultsmentioning

confidence: 99%

Identifying three-dimensional structures of autophosphorylation complexes in crystals of protein kinases

Malecka

Fink

et al. 2015

Sci. Signal.

View full text Add to dashboard Cite

show abstract

“…The equivalent region in Met was found to contain a mutation (S1058P) in a sample from the non small cell lung cancer [47], suggesting the potential functional significance of this region. In order to better understand the inhibitory mechanism imposed by the JM-C region, we compared the kinase domain sequences of 150 receptors by mapping them onto the crystal structure of the Ron kinase domain (3PLS) [48] (Figure 6). Based on surface accessibility and relative location, a conserved region on the surface of the Ron kinase N lobe was identified as a putative JM-C binding region (orange in Figure 6A).…”

Section: Discussionmentioning

confidence: 99%

“… Conserved surface residues and surface potential of the Ron kinase domain (3PLS) [ [48] ]. The conserved region adjacent to the active site with a predominantly positive surface potential is indicated by arrows as the putative JM-C binding site.…”

Section: Discussionmentioning

confidence: 99%

“…Based on the crystal structure of the Ron kinase domain, we built a molecular model of the inactive Ron structure (including JM-C, JM-D region and kinase domain) [48-51] (Additional file 3: Figure S3A), which suggests that the JM-C region (magenta) promoting a helical conformation could make close contacts with the P loop (orange) and activation loop (yellow). Acidic residues in the JM-C region (E 1044 -D 1045 -E 1046 ) have the potential to interact with H 1092 in the P-loop, Y 1238 and Q 1243 in the activation loop, and the side-chains of S 1117 and R 1118 between the β3 strand and the αC helix (lines in Additional file 3: Figure S3A).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Autoinhibition of the Ron receptor tyrosine kinase by the juxtamembrane domain

2014

View full text Add to dashboard Cite

BackgroundThe Ron receptor tyrosine kinase (RTK) has been implicated in the progression of a number of carcinomas, thus understanding the regulatory mechanisms governing its activity is of potential therapeutic significance. A critical role for the juxtamembrane domain in regulating RTK activity is emerging, however the mechanism by which this regulation occurs varies considerably from receptor to receptor.ResultsUnlike other RTKs described to date, tyrosines in the juxtamembrane domain of Ron are inconsequential for receptor activation. Rather, we have identified an acidic region in the juxtamembrane domain of Ron that plays a central role in promoting receptor autoinhibition. Furthermore, our studies demonstrate that phosphorylation of Y1198 in the kinase domain promotes Ron activation, likely by relieving the inhibitory constraints imposed by the juxtamembrane domain.ConclusionsTaken together, our experimental data and molecular modeling provide a better understanding of the mechanisms governing Ron activation, which will lay the groundwork for the development of novel therapeutic approaches for targeting Ron in human malignancies.

show abstract

“…RON is a transmembrane receptor with its short chain and N-terminal region of long chain presented on the cell surface, and the remaining part of the long chain containing tyrosine kinase and phosphorylation sites located intracellularly [23]. Binding of active MSP to RON triggers receptor autophosphorylation, leading to tyrosine kinase activation and consequent downstream signaling events [24,25].…”

Section: Msp and Its Receptor-ronmentioning

confidence: 99%