TY - JOUR
T1 - A pilot study on nitration/dysfunction of NK1 segment of myogenic stem cell activator HGF
AU - Elgaabari, Alaa
AU - Imatomi, Nana
AU - Kido, Hirochika
AU - Seki, Miyumi
AU - Tanaka, Sakiho
AU - Matsuyoshi, Yuji
AU - Nakashima, Takashi
AU - Sawano, Shoko
AU - Mizunoya, Wataru
AU - Suzuki, Takahiro
AU - Nakamura, Mako
AU - Anderson, Judy E.
AU - Tatsumi, Ryuichi
N1 - Funding Information:
To test the above hypothesis, we conducted preliminary in vitro experiments using peroxynitrite treatment of recombinant HGF and NK1 under physiological conditions. Results clearly demonstrated that functionally important Y198 residue in the K1 domain undergoes nitration and accounts for important results supporting that the nitration may abolish c-met binding and bromodeoxyuridine (BrdU)-incorporation activities of NK1. Physiological significance of this finding was supported by direct-immunofluorescence microscopy for rat lower hind-limb muscles, extensor digitorum longus (EDL) and tibialis anterior (TA) muscles from 2-month-old “young” and 12-month-old “retired/adult” groups; results showed that in vivo nitration of extracellular HGF (Y198) was detected more pronouncedly at 12-months old of the early aging-phase. Findings may provide biomedical strategies to combat dysfunction induced by HGF/NK1 nitration in age-related muscular disorders and diseases.This work was supported by KAKENHI Grant Numbers JP26660218 (Challenging Exploratory Research) and JP21H02347 (Scientific Research B) from the Japan Society for the Promotion of Science (to R.T.). A. Elgaabari was supported by the Egypt-Japan Education Partnership (EJEP), a full Ph.D. scholarship from the Egyptian Ministry of Higher Education. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript, and did not provide support in the form of salaries for any authors. Authors thank TORAY Co. for providing recombinant NK1. The anti-BrdU antibody (G3G4) developed by S. J. Kaufman and the anti-desmin antibody (D3) developed by D. A. Fischman were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biological Sciences, Iowa City, IA 52242, Iowa.
Funding Information:
This work was supported by KAKENHI Grant Numbers JP26660218 (Challenging Exploratory Research) and JP21H02347 (Scientific Research B) from the Japan Society for the Promotion of Science (to R.T.). A. Elgaabari was supported by the Egypt-Japan Education Partnership (EJEP), a full Ph.D. scholarship from the Egyptian Ministry of Higher Education . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript, and did not provide support in the form of salaries for any authors.
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/9
Y1 - 2022/9
N2 - Protein tyrosine residue (Y) nitration, a post-translational chemical-modification mode, has been associated with changes in protein activity and function; hence the accumulation of specific nitrated proteins in tissues may be used to monitor the onset and progression of pathological disorders. To verify the possible impact of nitration on postnatal muscle growth and regeneration, a pilot study was designed to examine the nitration/dysfunction of hepatocyte growth factor (HGF), a key ligand that is released from the extracellular tethering and activates myogenic stem satellite cells to enter the cell cycle upon muscle stretch and injury. Exposure of recombinant HGF (a hetero-dimer of α- and β-chains) to peroxynitrite induces Y nitration in HGF α-chain under physiological conditions. Physiological significance of this finding was emphasized by Western blotting that showed the NK1 segment of HGF (including a K1 domain critical for signaling-receptor c-met binding) undergoes nitration with a primary target of Y198. Peroxynitrite treatment abolished HGF-agonistic activity of the NK1 segment, as revealed by in vitro c-met binding and bromodeoxyuridine-incorporation assays. Importantly, direct-immunofluorescence microscopy of rat lower hind-limb muscles from two aged-groups (2-month-old “young” and 12-month-old “retired/adult”) provided in vivo evidence for age-related nitration of extracellular HGF (Y198). Overall, findings provide the insight that HGF/NK1 nitration/dysfunction perturbs myogenic stem cell dynamics and homeostasis; hence NK1 nitration may stimulate progression of muscular disorders and diseases including sarcopenia.
AB - Protein tyrosine residue (Y) nitration, a post-translational chemical-modification mode, has been associated with changes in protein activity and function; hence the accumulation of specific nitrated proteins in tissues may be used to monitor the onset and progression of pathological disorders. To verify the possible impact of nitration on postnatal muscle growth and regeneration, a pilot study was designed to examine the nitration/dysfunction of hepatocyte growth factor (HGF), a key ligand that is released from the extracellular tethering and activates myogenic stem satellite cells to enter the cell cycle upon muscle stretch and injury. Exposure of recombinant HGF (a hetero-dimer of α- and β-chains) to peroxynitrite induces Y nitration in HGF α-chain under physiological conditions. Physiological significance of this finding was emphasized by Western blotting that showed the NK1 segment of HGF (including a K1 domain critical for signaling-receptor c-met binding) undergoes nitration with a primary target of Y198. Peroxynitrite treatment abolished HGF-agonistic activity of the NK1 segment, as revealed by in vitro c-met binding and bromodeoxyuridine-incorporation assays. Importantly, direct-immunofluorescence microscopy of rat lower hind-limb muscles from two aged-groups (2-month-old “young” and 12-month-old “retired/adult”) provided in vivo evidence for age-related nitration of extracellular HGF (Y198). Overall, findings provide the insight that HGF/NK1 nitration/dysfunction perturbs myogenic stem cell dynamics and homeostasis; hence NK1 nitration may stimulate progression of muscular disorders and diseases including sarcopenia.
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U2 - 10.1016/j.bbrep.2022.101295
DO - 10.1016/j.bbrep.2022.101295
M3 - Article
AN - SCOPUS:85132314869
SN - 2405-5808
VL - 31
JO - Biochemistry and Biophysics Reports
JF - Biochemistry and Biophysics Reports
M1 - 101295
ER -