A750F Transmission A750F AUTOMATIC TRANSMISSION – AUTOMATIC TRANSMISSION UNIT AT–207 AT 16. 6) Fluid Type ATF Type T-IV ATF D-II or equivalent. No detectable His-tagged SAM domains could be observed in any of the GST-tagged Eph SAM domain pull-down experiments.A750f transmission pdf. The GSH-Sepharose beads pelleted proteins were separated by Tricine SDS-PAGE and stained with Coomassie blue. In this assay, each of purified GST-tagged Eph receptor SAM domain was incubated with a mixtures composed of the 14 His-tagged SAM domains (see 5% input lane). ( B) GST pull-down assay showing that none of the thirteen individual GST-tagged Eph receptor SAM domain had detectable binding to any one of the SAM domain in the 14 His-tagged SAM domain mixture. The fitted molar mass values of the His-SAM mixture, GST-SAM mixture, and the His-SAM + GST-SAM mixtures are also indicated in the figure, and further showing no detectable molar mass change upon mixing the two mixtures. The concentration of each SAM domain in the assay mixtures is 10 μM. the near perfect overlap of the blue, green and red elution profiles).
( A) SEC-MALS analyses showing that mixing the 14 His-tagged Eph receptor SAM domains with the 13 GST-tagged Eph receptor SAM domains (blue line) did not cause detectable elution profile changes when compared to the elution profile of the 14 His-tagged Eph receptor SAM domains (green line) and to that of 13 GST-tagged Eph receptor SAM domains (red line) (i.e. ( K) GST pull-down assay showing that the K857R mutant of EphA5 SAM gained its binding to SHIP2. ( J) The bindings of SHIP2 SAM or its F1226L-mutant to the full-length WT GFP-EphA2 analyzed by the GST pull-down assay. ( I) The Bindings of EphA2 SAM or its various mutants to the full-length SHIP2 by the GST pull-down assay. ( H) Summary of the ITC-derived results showing that the mutations of the critical residues in the SAM domain interface can dramatically affect the bindings of EphA SAMs to the SAM domains from SHIP2 or Odin. Note the planar alignment of the π electrons of the Arg sidechain and the neighboring peptide backbones in both structures. ( F–G) Details of the planar cation-π interaction between EphA2 R958 and SHIP2 F1226 ( F), and EphA6 R1108 and Odin F743 ( G). ( E) Superposition of two the EphA2 SAM-SHIP2 SAM and the EphA6 SAM-Odin SAM1 complex structures with the critical R958/F1226 and R1108/F743 pairs highlighted with the stick model. The Gly-aromatic residue pairs (Gly954 A2/Gly1104 A6 and Trp1221 SHIP2/Phe738 Odin) are indicated with red dashed circles. Salt bridges and hydrogen bonds are indicated with dashed lines. Key residues are shown with the stick model. ( B and D) Details of the interfaces in the EphA2 SAM-SHIP2 SAM ( B) and EphA6 SAM-Odin SAM1 ( D) complexes.
( A and C) Ribbon representations of the EphA2 SAM-SHIP2 SAM ( A) and the EphA6 SAM-Odin SAM1 ( C) complex structures.
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