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USP4 binds Nmdar1. 24 / 24
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"It has a kinase domain near the C-terminus that is homologous to the catalytic domain of MLCK 1 Abbreviations used: NMR, unique N-terminal domain of myorod; NMR-unP and NMR-P, synthetic unphosphorylat[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"AJ249993 , [7] ) were chemically synthesized by ShineGene Molecular Biotech (Shanghai, China) with phosphorylated Thr141 (NMR-P) and a substitution of Thr residue for Ala141 (NMR-unP) to represent the[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"This peptide corresponding to the unique N-terminal region of the amino acid sequence of myorod was synthesized either in phosphorylated (NMR-P) or unphosphorylated (NMR-unP) form (see Materials and m[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"The concentration of myosin held constant and increasing amount of NMR-P and NMR-unP were used in a co-sedimentation assay to determine the binding affinity for the peptides."
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"We found that the peptides were able to bind to myosin and the binding affinity of NMR-P to myosin was higher ( K app = 0.055 μM) than that of NMR-unP ( K app = 0.116 μM)."
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"Only NMR-unP peptide caused the sedimentation of F-actin; the amount of F-actin detected in low-speed pellets increased with increasing concentrations of NMR-unP peptide in the assay mixture ( Fig. 2 [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"NMR-unP can also interact with molluscan thin filaments."
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"However, we note a distinctive feature of this binding: the NMR-unP peptide co-sedimentes with thin filaments only in the presence of Ca 2+ ( Fig. 2 B, lanes 3–6), while interaction of NMR-unP with F-[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"As shown in Fig. 3 , the viscosity of F-actin decreased in the presence of NMR-unP in a concentration-dependent manner, indicating that aggregates or bundles of F-actin have formed."
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"To explore the bundling activity of the NMR-unP peptide further, we analyzed the structural arrangement of fluorescently labeled actin filaments after incubation with increasing concentrations of NMR-[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"As shown in Fig. 4 , NMR-unP caused the formation of dense actin bundles."
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"At low NMR-unP concentrations, it is more probable that loose network is formed ( Fig. 4 C)."
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"As the concentration of NMR-unP is increased, more tightly bound individual bundles become visible in an otherwise largely homogeneous network of actin filaments ( Fig. 4 D–F)."
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"We noted that the network formation ( Fig. 4 E) was sufficient to completely sediment the F-actin or thin filaments in the presence of NMR-unP by low-speed centrifugation ( Fig. 2 )."
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"As shown above, the interaction of the NMR-unP peptide with F-actin caused a significant decrease in the viscosity of the actin filaments ( Fig. 3 )."
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"Thus, full-length myorod and its NMR-unP peptide binds to thin filaments in a Ca 2+ -sensitive manner, while NMR-unP–F-actin interaction is Ca 2+ -independent ( Fig. 2 C)."
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"Taken together, these results indicate that full-length myorod is able to interact with filamentous actin and thin filaments with the same bundling activity and influence on actin viscosity that was o[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"We found that NMR-unP peptide of myorod binds to rabbit F-actin in a Ca 2+ -independent manner and to molluscan thin filaments in a Ca 2+ -dependent manner."
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"The aggregates were visualized by fluorescence microscopy, which revealed the bundles of actin filaments created in the presence of NMR-unP ( Fig. 4 )."
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"The formation of myorod–actin complexes caused a fall in the viscosity and the appearance of bundles of actin filaments visualized by fluorescence microscopy as for the formation of NMR-unP peptide–ac[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"The mechanism by which NMR-unP peptide and full-length myorod mediate F-actin bundling activity remains unclear."
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"As mentioned earlier, the interaction of NMR-unP peptide and full-length myorod with thin filaments was Ca 2+ -dependent; the interaction was inhibited in the absence of Ca 2+ ."
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"Thus, the inhibition of the NMR-unP peptide or full-length myorod interaction with thin filaments in the absence of Ca 2+ suggests that the myorod binding sites on actin are, supposedly, occupied with[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Importantly, only NMR-unP and unphosphorylated full-length protein are able to interact with filamentous actin and thin filaments, unlike NMR-P which, in turn, has higher affinity for myosin in compar[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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