Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle.
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 novel 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 lysylendopeptidase, 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.
Hayashi
T
T
Laboratory of Chemistry and Technology of Animal Products, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
Takeshita
K
K
Tsuchida
N
N
Kitano
K
K
Kawabata
S
S
Iwanaga
S
S
Ito
T
T
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000302:isolation & purification*; D036341:Intercellular Signaling Peptides and Proteins; D018482:Muscle, Skeletal / Q000737:chemistry*; D011817:Rabbits; D013552:Swine
D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; C065700:myotrophin