title: Ankyrin domain of myosin 16 influences motor function and decreases protein phosphatase catalytic activity.
abstract: The unconventional myosin 16 (Myo16), which may have a role in regulation of cell cycle and cell proliferation, can be found in both the nucleus and the cytoplasm. It has a unique, eight ankyrin repeat containing pre-motor domain, the so-called ankyrin domain (My16Ank). Ankyrin repeats are present in several other proteins, e.g., in the regulatory subunit (MYPT1) of the myosin phosphatase holoenzyme, which binds to the protein phosphatase-1 catalytic subunit (PP1c). My16Ank shows sequence similarity to MYPT1. In this work, the interactions of recombinant and isolated My16Ank were examined in vitro. To test the effects of My16Ank on myosin motor function, we used skeletal muscle myosin or nonmuscle myosin 2B. The results showed that My16Ank bound to skeletal muscle myosin (K D ≈ 2.4 µM) and the actin-activated ATPase activity of heavy meromyosin (HMM) was increased in the presence of My16Ank, suggesting that the ankyrin domain can modulate myosin motor activity. My16Ank showed no direct interaction with either globular or filamentous actin. We found, using a surface plasmon resonance-based binding technique, that My16Ank bound to PP1cα (K D ≈ 540 nM) and also to PP1cδ (K D ≈ 600 nM) and decreased its phosphatase activity towards the phosphorylated myosin regulatory light chain. Our results suggest that one function of the ankyrin domain is probably to regulate the function of Myo16. It may influence the motor activity, and in complex with the PP1c isoforms, it can play an important role in the targeted dephosphorylation of certain, as yet unidentified, intracellular proteins.
authors.affiliation: Department of Biophysics, University of Pécs Medical School, Szigeti Str. 12, Pécs, H-7624, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Kengyel
authors.forename: Bálint
authors.initials: B
authors.lastname: Bécsi
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Kónya
authors.forename: James R
authors.initials: JR
authors.lastname: Sellers
authors.forename: Ferenc
authors.initials: F
authors.lastname: Erdődi
authors.forename: Miklós
authors.initials: M
authors.lastname: Nyitrai
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D017089:Ankyrin Repeat; D011485:Protein Binding; D054645:Protein Phosphatase 1 / Q000378:metabolism*; D051381:Rats; D024744:Skeletal Muscle Myosins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000199:Actins; D054645:Protein Phosphatase 1; D024744:Skeletal Muscle Myosins