Department of Biochemistry, University of Umeå, S-901 87 Umeå, Sweden.
Plant Physiol. 1980 Sep;66(3):442-5. doi: 10.1104/pp.66.3.442.
Mitochondria from green leaves of spinach have been prepared using a three-step procedure involving differential centrifugation, partition in an aqueous dextran polyethylene glycol two-phase system and Percoll gradient centrifugation. The mitochondrial fractions after the different steps of purification were compared. The final mitochondrial preparation was totally free from chloroplast material measured as chlorophyll content. The enrichment of mitochondria in relation to peroxisomes and microsomes was approximately 12 and 33 times, respectively, based on NAD:isocitrate dehydrogenase activity, glycolate oxidase activity, and NADPH:cytochrome c oxidoreductase activity. The apparent intactness of the inner and the outer mitochondrial membranes was higher than 90% as measured by latency of enzyme activities. The mitochondria showed high respiratory rates with respiratory control and the ADP/O ratios approached the theoretical limits.
菠菜的绿色叶片中的线粒体,已通过涉及差速离心、在水-葡聚糖聚乙二醇两相系统和Percoll 梯度离心中的分配的三步程序来制备。对不同纯化步骤的线粒体部分进行了比较。最终的线粒体制剂完全不含叶绿体物质,这可通过叶绿素含量来衡量。基于 NAD:异柠檬酸脱氢酶活性、乙醇酸氧化酶活性和 NADPH:细胞色素 c 氧化还原酶活性,线粒体相对于过氧化物酶体和微粒体的富集程度分别约为 12 倍和 33 倍。酶活性的潜伏性测量表明,内外线粒体膜的完整性明显高于 90%。线粒体表现出高呼吸速率和呼吸控制,ADP/O 比接近理论极限。