Myosin as a potential redox-sensor: an in vitro study.
A balanced redox status is necessary to optimize force production in contractile apparatus, where free radicals generated by skeletal muscle are involved in some basic physiological processes like excitation-contraction coupling. Protein glutathionylation has a key role in redox regulation of proteins and signal transduction. Here we show that myosin is sensitive to in vitro glutathionylation and MALDI-TOF analysis identified three potential sites of glutathione binding, two of them locating on the myosin head. Glutathionylation of myosin has an important impact on the protein structure, as documented by the lower fluorescence quantum yield of glutathionylated myosin and its increased susceptibility to the proteolytic cleavage. Myosin function is also sensitive to glutathionylation, which modulates its ATPase activity depending on GSSG redox balance. Thus, like the phosphorylation/dephosphorylation cycle, glutathionylation may represent a mechanism by which glutathione modulates sarcomere functions depending on the tissue redox state, and myosin may constitute a muscle redox-sensor.
Passarelli
Chiara
C
Molecular Medicine Unit, Children's Hospital and Research Institute Bambino Gesù, Department of Biology, University of Rome Roma Tre, P.za S. Onofrio, 4, Rome, 00165, Italy.
Petrini
Stefania
S
Pastore
Anna
A
Bonetto
Valentina
V
Sale
Patrizio
P
Gaeta
Laura M
LM
Tozzi
Giulia
G
Bertini
Enrico
E
Canepari
Monica
M
Rossi
Rosetta
R
Piemonte
Fiorella
F
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002384:Catalysis / Q000187:drug effects; D004229:Dithiothreitol / Q000737:chemistry / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry* / Q000378:metabolism; D019803:Glutathione Disulfide / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D009218:Myosins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; D014357:Trypsin; D000251:Adenosine Triphosphatases; D009218:Myosins; D005978:Glutathione; D004229:Dithiothreitol; D019803:Glutathione Disulfide
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