QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. Interaction of the N-terminal region of obscurin/Obsl1 with myomesin. (A) Sub-mapping of the minimal binding domain of obscurin/Obsl1 to My4-My5 and of myomesin to obscurin/Obsl1 Ig3 by successive deletion mutants of My2-My8 using Y2H assays identified Ig3 of obscurin/Obsl1 (top) and the linker between My4 and My5 of myomesin (bottom) as sufficient for binding. (B) These interactions were verified by pulldown assays using GST–My4-My5-linker bound to glutathione beads and GFP-tagged obscurin/Obsl1 Ig3 constructs. (B,C) Detection was performed by blotting equivalent amounts of input (I), unbound (U) and bound (P) fractions with anti-GFP antibody. (C) The interaction of obscurin/Obsl1 Ig3 with the linker between myomesin My4 and My5 is specific for myomesin and is not regulated by phosphorylation. (C, top) Predicted secondary structure of the My4-My5 linker with the protein kinase A (PKA) site. (C, middle) Alignment of the linker sequence of the three myomesin gene family members, MYOM1-MYOM3, with the phosphorylated serine is shown by a black arrow, the neighbouring serine 615 is shown by the grey arrow; red, residues conserved in all three genes; blue, residues conserved only in two MYOM genes. (C, bottom) Pulldown assay with mutation of the phosphorylated serine 618 in MYOM1 to alanine (SA) or aspartate (SD).

Return to article