Borjigin Gerelt, Yamamoto Shuhei, Sugiyama Toshie, Nishiumi Tadayuki, Suzuki Atsushi
Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Japan.
Biosci Biotechnol Biochem. 2006 May;70(5):1249-54. doi: 10.1271/bbb.70.1249.
Changes in the localization of calpain in conditioned and pressurized bovine skeletal muscles were investigated by immunogold electron-microscopy. In the muscle immediately after thawing (control), the relative distributions of colloidal particles statistically calculated by counting the colloidal particles were about 65% and 35% in the I-band/Z-disk and A-band regions, respectively. In the muscle conditioned for 7 days, distribution of colloidal particles was more than two times greater in both the I-band/Z-disk and A-band regions than in the control muscle. Almost no change in either the absolute concentration or relative distribution of the colloidal particles was detectable during further storage. In the muscle exposed to a pressure of 100 MPa or 200 MPa, slightly more immunogold was detected in the sarcomere than in that of the control muscle. Increasing pressure up to 300 MPa enabled high-density particles to be seen throughout the sarcomere. Conversely, few particles were detected in the sarcomere of the muscle exposed to 400 MPa. These electron-microscopic observations were confirmed from the statistical analysis as with the conditioned muscles. It was clear from the results obtained that the pressure-induced changes in calpain localization were much more pronounced than those from conditioning.
通过免疫金电子显微镜研究了钙蛋白酶在条件处理和加压的牛骨骼肌中的定位变化。在解冻后立即的肌肉(对照)中,通过计数胶体颗粒统计计算的胶体颗粒相对分布在I带/Z盘和A带区域分别约为65%和35%。在条件处理7天的肌肉中,I带/Z盘和A带区域的胶体颗粒分布比对照肌肉中的多两倍以上。在进一步储存期间,胶体颗粒的绝对浓度或相对分布几乎没有变化。在暴露于100MPa或200MPa压力的肌肉中,在肌节中检测到的免疫金略多于对照肌肉。将压力增加到300MPa可使高密度颗粒在整个肌节中可见。相反,在暴露于400MPa的肌肉的肌节中几乎检测不到颗粒。这些电子显微镜观察结果通过与条件处理肌肉相同的统计分析得到证实。从获得的结果可以清楚地看出,压力诱导的钙蛋白酶定位变化比条件处理引起的变化更为明显。