Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle

Toshiya Hayashi, Keisuke Takeshita, Nobuyuki Tsuchida, Kiyokazu Kitano, Shun-Ichiro Kawabata, Sadaaki Iwanaga, Tatsumi Ito

Research output: Contribution to journalArticle

6 Citations (Scopus)

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 languageEnglish
Pages (from-to)897-908
Number of pages12
JournalInternational Journal of Biochemistry and Cell Biology
Volume30
Issue number8
DOIs
Publication statusPublished - Aug 1 1998

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Cell growth
Muscle Cells
Purification
Muscle
Intercellular Signaling Peptides and Proteins
Skeletal Muscle
Swine
lysyl endopeptidase
Water
Muscle Denervation
Thermolysin
Muscles
Periodic Acid
Inbred SHR Rats
Growth
Cells
Polyacrylamide Gel Electrophoresis
Digestion
Leg
Myocardium

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology

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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 journalArticle

Hayashi, Toshiya ; Takeshita, Keisuke ; Tsuchida, Nobuyuki ; Kitano, Kiyokazu ; Kawabata, Shun-Ichiro ; Iwanaga, Sadaaki ; Ito, Tatsumi. / Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle. In: International Journal of Biochemistry and Cell Biology. 1998 ; Vol. 30, No. 8. pp. 897-908.
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