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  • Title: Isoenzyme-specific localization of M-line bound creatine kinase in myogenic cells.
    Author: Wallimann T, Moser H, Eppenberger HM.
    Journal: J Muscle Res Cell Motil; 1983 Aug; 4(4):429-41. PubMed ID: 6355174.
    Abstract:
    Experiments using isolated fibre bundles or myofibrils of chicken skeletal muscle have shown that a relatively small portion of the muscle-specific MM-type of creatine kinase (CK) (EC 2.7.3.2) is specifically bound to the M-line and yet greatly contributes to the electron-dense M-line structure. Here we demonstrate the presence of M-line bound CK in cultured myogenic cells by removing the unbound sarcoplasmic CK through permeabilization with Triton X-100 and extensive washing of the cells prior to immunofluorescence staining. When stained with antibodies specific for M-CK subunits these cells exhibit bright fluorescence within the M-line region of myofibrils. Occasionally this cross-striated pattern is also observed in mononucleated presumably postmitotic myoblasts. Anti-B-CK incubation, in contrast, results in a weak, diffuse fluorescence at the Z-band. Even though these cells contain appreciable amounts of B-type CK, specific fluorescence at the M-line is never observed with anti-B-CK antibody thus ruling out the presence of BB-type or MB-type CK at this location. Therefore the presence of CK within the M-line structure of myogenic cells which contain all three CK isoenzymes seems to be restricted to the MM-type isoenzyme.
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