Abstract
A comparison of muscle weight between denerved and control rabbit hind legs revealed that a water-soluble 12 kDa substance was reduced in atrophied muscles after denervation. We hypothesised that a water-soluble growth factor exists which mediates a signal from motor nerves to muscles. To isolate this factor we modified the purification procedures of Sen et al. [S. Sen, G. Kundu, N. Mekhail, J. Castel, K. Misono, B. Healy, Myotrophin: purification of a navel peptide from spontaneously hypertensive rat heart that influences myocardial growth, J. Biol. Chem, 265 (1990) 16635-16643.], who originally purified a water-soluble growth factor from cardiac muscles. Four additional purification steps were added to the method. Using this technique, a novel muscle cell growth factor, named s-myotrophin, was purified from porcine skeletal muscle (M. longissimus thoracis). Purified s-myotrophin appeared as a single band (12 kDa) on SDS-PAGE and had a strong growth promoting activity (increase of protein synthesis) of cultured primary skeletal muscle cells. Almost no loss of growth promoting activity was observed after trypsin and chymotrypsin digestion, No fragmentation of s-myotrophin was observed after exposure to lysyl-endopeptidase, thermolysin, trypsin and chymotrypsin. Crude preparation of this molecule could be detected by periodic acid/Schiff (PAS) staining. Deglycosylation of S-myotrophin produced a smaller molecule having an approximately 7 kDa mass. These data indicate a novel 12 kDa protein has been isolated which has growth promoting properties on skeletal muscle cells.
| Original language | English |
|---|---|
| Pages (from-to) | 897-908 |
| Number of pages | 12 |
| Journal | International Journal of Biochemistry and Cell Biology |
| Volume | 30 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 1 1998 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Cell Biology
Cite this
Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle. / Hayashi, Toshiya; Takeshita, Keisuke; Tsuchida, Nobuyuki; Kitano, Kiyokazu; Kawabata, Shun-Ichiro; Iwanaga, Sadaaki; Ito, Tatsumi.
In: International Journal of Biochemistry and Cell Biology, Vol. 30, No. 8, 01.08.1998, p. 897-908.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle
AU - Hayashi, Toshiya
AU - Takeshita, Keisuke
AU - Tsuchida, Nobuyuki
AU - Kitano, Kiyokazu
AU - Kawabata, Shun-Ichiro
AU - Iwanaga, Sadaaki
AU - Ito, Tatsumi
PY - 1998/8/1
Y1 - 1998/8/1
N2 - A comparison of muscle weight between denerved and control rabbit hind legs revealed that a water-soluble 12 kDa substance was reduced in atrophied muscles after denervation. We hypothesised that a water-soluble growth factor exists which mediates a signal from motor nerves to muscles. To isolate this factor we modified the purification procedures of Sen et al. [S. Sen, G. Kundu, N. Mekhail, J. Castel, K. Misono, B. Healy, Myotrophin: purification of a navel peptide from spontaneously hypertensive rat heart that influences myocardial growth, J. Biol. Chem, 265 (1990) 16635-16643.], who originally purified a water-soluble growth factor from cardiac muscles. Four additional purification steps were added to the method. Using this technique, a novel muscle cell growth factor, named s-myotrophin, was purified from porcine skeletal muscle (M. longissimus thoracis). Purified s-myotrophin appeared as a single band (12 kDa) on SDS-PAGE and had a strong growth promoting activity (increase of protein synthesis) of cultured primary skeletal muscle cells. Almost no loss of growth promoting activity was observed after trypsin and chymotrypsin digestion, No fragmentation of s-myotrophin was observed after exposure to lysyl-endopeptidase, thermolysin, trypsin and chymotrypsin. Crude preparation of this molecule could be detected by periodic acid/Schiff (PAS) staining. Deglycosylation of S-myotrophin produced a smaller molecule having an approximately 7 kDa mass. These data indicate a novel 12 kDa protein has been isolated which has growth promoting properties on skeletal muscle cells.
AB - A comparison of muscle weight between denerved and control rabbit hind legs revealed that a water-soluble 12 kDa substance was reduced in atrophied muscles after denervation. We hypothesised that a water-soluble growth factor exists which mediates a signal from motor nerves to muscles. To isolate this factor we modified the purification procedures of Sen et al. [S. Sen, G. Kundu, N. Mekhail, J. Castel, K. Misono, B. Healy, Myotrophin: purification of a navel peptide from spontaneously hypertensive rat heart that influences myocardial growth, J. Biol. Chem, 265 (1990) 16635-16643.], who originally purified a water-soluble growth factor from cardiac muscles. Four additional purification steps were added to the method. Using this technique, a novel muscle cell growth factor, named s-myotrophin, was purified from porcine skeletal muscle (M. longissimus thoracis). Purified s-myotrophin appeared as a single band (12 kDa) on SDS-PAGE and had a strong growth promoting activity (increase of protein synthesis) of cultured primary skeletal muscle cells. Almost no loss of growth promoting activity was observed after trypsin and chymotrypsin digestion, No fragmentation of s-myotrophin was observed after exposure to lysyl-endopeptidase, thermolysin, trypsin and chymotrypsin. Crude preparation of this molecule could be detected by periodic acid/Schiff (PAS) staining. Deglycosylation of S-myotrophin produced a smaller molecule having an approximately 7 kDa mass. These data indicate a novel 12 kDa protein has been isolated which has growth promoting properties on skeletal muscle cells.
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UR - http://www.scopus.com/inward/citedby.url?scp=0032146296&partnerID=8YFLogxK
U2 - 10.1016/S1357-2725(98)00052-1
DO - 10.1016/S1357-2725(98)00052-1
M3 - Article
C2 - 9744081
AN - SCOPUS:0032146296
VL - 30
SP - 897
EP - 908
JO - International Journal of Biochemistry and Cell Biology
JF - International Journal of Biochemistry and Cell Biology
SN - 1357-2725
IS - 8
ER -