Distribution of cathepsins B, H, L, and trypsin-like proteases in natural actomyosin from washed meat of various fishes

Hu, Yaqin; Morioka, Katsuji; Itoh, Yoshiaki

doi:10.1111/j.1444-2906.2008.01577.x

Cited by 10 publications

(16 citation statements)

References 11 publications

(16 reference statements)

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“…The total activity of cathepsin L‐like that remained in AM samples for spotted chub mackerel was the highest, intermediate activity for true lizardfish, and the lowest activity for redbanded searobin. The tendency was consistent with those activities detected in muscles for the respective fish species 1 . The result also coincided with the report that cathepsin activities in red‐flesh fishes were higher than that in white‐flesh fishes 7 .…”

supporting

confidence: 92%

“…Specific activities of rat liver cathepsin L and B on Z‐Phe‐Arg‐MCA were reported to be 1229 and 400 U/μmol enzyme, respectively, while those on Z‐Arg‐Arg‐MCA were 2.4 and 367 U/μmol, respectively 6 . In a previous report, 1 after gel filtration, profiles of cathepsin B‐like in the AM from the 18 species of tested materials were quite different from that of cathepsin L‐like, and the activity value of cathepsin B‐like was much lower than that of cathepsin L‐like. Therefore, the cathepsin B‐like activity could be negligible when Z‐Phe‐Arg‐MCA was used as a substrate.…”

mentioning

confidence: 93%

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Effect of meat-bleaching and dilution-precipitation procedures on the removal of cathepsin L-like contained in the actomyosin of various fish species

Morioka

Itoh

2008

Fisheries Sci

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KEY WORDS: actomyosin, bleaching, cathepsin L-like, dilution-precipitation, redbanded searobin, spotted chub mackerel, true lizardfish.In a previous report, it was found that cathepsins B, H, L and trypsin-like proteases still remained in the natural actomyosin (AM). 1 Cathepsin L-like was found to be present as a detached form from AM, as studied by gel filtration, despite that the activity could not be removed by repeated bleaching of meat. It was also found that the activity varied from fish species to species. Hu et al. found that spotted chub mackerel, true lizardfish and redbanded searobin showed the highest, middle and the lowest cathepsin L-like activities, respectively. 1 Actomyosin used in the study was prepared from fish mince bleached three times, and purified by a cycle of dilution-precipitation procedures. 1 It might be necessary to investigate if the above treatments would affect the remaining activity of cathepsin L-like. The objective of this present study was to investigate if the activity differences among fish species of cathepsin L-like were affected by bleaching and dilution-precipitation.Spotted chub mackerel Scomber australasicus (38.5 cm, 729.0 g), true lizardfish Saurida undosquamis (three fishes with mean fork length of 35.7 cm, mean body weight of 159.8 g) and redbanded searobin Lepidotrigla guentheri (seven fishes with mean fork length of 14.7 cm, mean body weight of 42.3 g) were used as materials. The fresh fishes were purchased from Kochi Central Fish Market (Kochi prefecture, Japan) and were brought to the laboratory in ice and were used immediately.Actomyosin was prepared according to the method of Takashi and Arai. 2 All of the procedures were performed at 4°C unless otherwise indicated. Approximately 40 g of dorsal muscle was homogenized with 4 volumes of I = 0.05 phosphate buffer (buffer A, pH 7.5) using a non-bubbling homogenizer (Diameter 3.6 cm; Nihonseiki Kaisha Ltd, Tokyo Japan) and centrifuged at 3000 ¥g for 10 min (Hitachi CR21E; Hitachi, Tokyo, Japan). This treatment was defined as a single bleaching of mince. The same procedures were repeated three times. The resulting supernatant by the procedures was referred to as SP with the numbers of bleaching. The pellet at the stage was collected and was dispersed in buffer A containing 0.6 M NaCl (buffer B). After standing at 4°C for 20 h, the sample was centrifuged at 10 000 ¥g for 30 min to obtain AM in the supernatant which was referred as AM with the numbers of bleaching.The NaCl concentration for SP fractions was adjusted to 0.6 M by adding solid NaCl.In order to purify AM, a dilution-precipitation procedure was applied. AM3 was used as a starting material. AM3 was diluted with 10 volumes of cold deionized water and centrifuged at 3000 ¥g for 10 min. This was defined as a single cycle of dilution-precipitation procedures. The procedures were repeated three times. The supernatant and the AM pellet were termed as DP and AM3 with the numbers of treatment. AM and DP fractions were both dissolved into 0.6 M NaCl.As a control, ...

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confidence: 92%

mentioning

confidence: 93%

Effect of meat-bleaching and dilution-precipitation procedures on the removal of cathepsin L-like contained in the actomyosin of various fish species

Morioka

Itoh

2008

Fisheries Sci

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“…In a previous study, cathepsin L was confirmed to remain in the actomyosin despite intensive leaching; after gel filtration of actomyosin the main activity peak of cathepsin L was clearly separated from that of actomyosin and was revealed to be an actomyosin non-binding cathepsin L. 14 Fractions showing the www.interscience.wiley.com/jsfa main activity peak of cathepsin L were pooled to obtain partially purified cathepsin L (L mix , Fig. 3).…”

Section: Preparation Of Partially Purified Cathepsin L (L Mix )mentioning

confidence: 99%

“…Actomyosin was prepared using the method of Hu et al 14 The surimi was first washed three times with low-ionic-strength buffer and then subjected to one cycle of dilution/precipitation. The sample of actomyosin obtained was resuspended in 0.6 mol L −1 NaCl/50 mmol L −1 phosphate buffer (pH 7.0) in preparation for gel filtration.…”

Section: Sample Preparationmentioning

confidence: 99%

Participation of cysteine protease cathepsin L in the gel disintegration of red bulleye (Priacanthus macracanthus) surimi gel paste

Morioka

Itoh

2009

J Sci Food Agric

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“…Sub-cellular compartments are disrupted, enzyme-inhibitor complexes may be formed or dissociated and enzyme precursors may be activated. It is well known that myofibrillar proteins (surimi) obtained after homogenisation and washing of fish muscle possess proteolytic enzyme activities (An, Weerasinghe, Seymour, & Morrissey, 1994;Cao, Jiang, Zhong, Zhang, & Su, 2006;Hu, Morioka, & Itoh, 2008;Wang, Martinez, & Olsen, 2009). It is however an open question if these activities are native to the myofibrillar proteins or if enzyme-myofibrillar complexes are formed during the preparation of the myofibrillar fraction.…”

Section: Introductionmentioning

confidence: 99%

Post-mortem degradation of myosin heavy chain in intact fish muscle: Effects of pH and enzyme inhibitors

et al. 2011

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a b s t r a c tFish muscle is rapidly degraded during post-mortem storage, due to proteolytic enzymes acting probably both on muscle cells and connective tissue. In this work we have developed a model system which may be used to study the enzymatic degradation occurring in intact post-mortem fish muscle. Degradation of myosin heavy chain (MHC) was monitored in muscle with pH adjusted to 6.05, 6.3 and 6.9 and in the presence of the enzyme inhibitors PMSF, EDTA, phenanthroline, pepstatin A, antipain, E-64 and the cysteine proteinase activator dithiothreithol (DTT). After storage, myofibrillar proteins were isolated and MHC-specific antibodies used to study the degradation in the different samples. MHC from muscle with pH 6.05 and 6.3 was degraded, while no severe degradation was observed at pH 6.9. Introduction of enzyme inhibitors into the muscle tissue clearly showed that mainly cysteine and aspartic proteinases are responsible for the in situ MHC degradation. This is supported by the severe breakdown of MHC in the muscle samples containing DTT.

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Distribution of cathepsins B, H, L, and trypsin-like proteases in natural actomyosin from washed meat of various fishes

Cited by 10 publications

References 11 publications

Effect of meat-bleaching and dilution-precipitation procedures on the removal of cathepsin L-like contained in the actomyosin of various fish species

Effect of meat-bleaching and dilution-precipitation procedures on the removal of cathepsin L-like contained in the actomyosin of various fish species

Participation of cysteine protease cathepsin L in the gel disintegration of red bulleye (Priacanthus macracanthus) surimi gel paste

Post-mortem degradation of myosin heavy chain in intact fish muscle: Effects of pH and enzyme inhibitors

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