Occurrence of Gelatinolytic Activities in Yellowtail Tissues

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  • Kubota Satoshi
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University
  • Toyohara Haruhiko
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University
  • Sakaguchi Morihiko
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University

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  • Occurrence of Gelatinolytic Activities

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Various gelatinolytic activities in phosphate-buffered saline extracts of yellowtail skin, muscle, gill, heart, liver, spleen, stomach, pyloric caeca, intestine, and caudal fin were revealed by gelatin zymographic analysis. Most active bands were demonstrated at larger than 50 kDa in all tissues tested, but the additional active bands smaller than 50 kDa were also detected in the pyloric caeca. An active band at 80 kDa sensitive to leupeptin (acetyl-L-leucyl-L-leucyl-L-argininal) was commonly detected in all tissues. Active bands sensitive to leupeptin at 140 kDa and 80-100 kDa in the gill and at 110 kDa in the pyloric caeca and at 120 kDa in the intestine were distributed specifically in these tissues. Intensive active bands between 25-80 kDa in the pyloric caeca were also sensitive to leupeptin. However, these active bands sensitive to leupeptin were not affected by E-64 ([L-3-trans-carboxyoxirane-2-carbonyl]-L-leucyl-agmatin) except for those at 110 kDa in the pyloric caeca and at 120 kDa in the intestine, suggesting that most active bands sensitive to leupeptin were derived from serine-proteinases. Active bands sensitive to 1, 10-phenanthroline were detected in the following tissues: skin, muscle, heart, liver, spleen, stomach, and caudal fin at 80 kDa, all tissues at 60 and 55 kDa, in the skin, gill, spleen, and caudal fin at 85 kDa, spleen at 70 kDa, and skin, gill, and caudal fin at 65 kDa, suggesting gelatinolytic metalloproteinase activities were widely distributed in yellowtail tissues. p-AminophenyImercuric acetate induced various latent serine-proteinase activities only in the gill.

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