Interface analysis of the complex between ERK2 and PTP-SL.
The activity of ERK2, an essential component of MAP-kinase pathway, is under the strict control of various effector proteins. Despite numerous efforts, no crystal structure of ERK2 complexed with such partners has been obtained so far. PTP-SL is a major regulator of ERK2 activity. To investigate the ERK2-PTP-SL complex we used a combined method based on cross-linking, MALDI-TOF analysis, isothermal titration calorimetry, molecular modeling and docking. Hence, new insights into the stoichiometry, thermodynamics and interacting regions of the complex are obtained and a structural model of ERK2-PTP-SL complex in a state consistent with PTP-SL phosphatase activity is developed incorporating all the experimental constraints available at hand to date. According to this model, part of the N-terminal region of PTP-SL has propensity for intrinsic disorder and becomes structured within the complex with ERK2. The proposed model accounts for the structural basis of several experimental findings such as the complex-dissociating effect of ATP, or PTP-SL blocking effect on the ERK2 export to the nucleus. A general observation emerging from this model is that regions involved in substrate binding in PTP-SL and ERK2, respectively are interacting within the interface of the complex.
Department of Enzymology, Institute of Biochemistry, Bucharest, Romania.
Mihaela C
MC
Balasu
Laurentiu N
LN
Spiridon
Simona
S
Miron
Constantin T
CT
Craescu
Axel J
AJ
Scheidig
Andrei-Jose
AJ
Petrescu
Stefan E
SE
Szedlacsek
D000595:Amino Acid Sequence; D002151:Calorimetry; D002850:Chromatography, Gel; D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding / Q000187:drug effects; D054634:Receptor-Like Protein Tyrosine Phosphatases, Class 7 / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003432:Cross-Linking Reagents; D010455:Peptides; D019950:Mitogen-Activated Protein Kinase 1; D054634:Receptor-Like Protein Tyrosine Phosphatases, Class 7