Hulbert A J, Turner Nigel, Hinde Jack, Else Paul, Guderley Helga
Metabolic Research Centre and School of Biological Sciences, University of Wollongong, Wollongong, NSW, 2522, Australia.
J Comp Physiol B. 2006 Feb;176(2):93-105. doi: 10.1007/s00360-005-0025-z. Epub 2006 Jan 12.
Mitochondria were isolated from the liver, kidney and mixed hindlimb skeletal muscle of three vertebrate species; the laboratory rat Rattus norvegicus, the bearded dragon lizard Pogona vitticeps, and the cane toad Bufo marinus. These vertebrate species are approximately the same body mass and have similar body temperatures. The content of cytochromes B, C, C1, and A were measured in these isolated mitochondria by oxidised-reduced difference spectra. Adenine nucleotide translocase (ANT) was measured by titration of mitochondrial respiration with carboxyactractyloside and the protein and phospholipid content of isolated mitochondria were also measured. Fatty acid composition of mitochondrial phospholipids was measured. Mitochondrial respiration was measured at 37 degrees C under states III and IV conditions as well as during oligomycin inhibition. Species differed in the ratios of different mitochondrial cytochromes. Muscle mitochondria differed from kidney and liver mitochondria by having a higher ANT content relative to cytochrome content. Respiration rates were compared relative to a number of denominators and found to be most variable when expressed relative to mitochondrial protein content and least variable when expressed relative to mitochondrial cytochrome A and ANT content. The turnover of cytochromes was calculated and found to vary between 1 and 94 electrons s(-1). The molecular activity of mitochondrial cytochromes was found to be significantly positively correlated with the relative polyunsaturation of mitochondrial membrane lipids.
从三种脊椎动物的肝脏、肾脏和后肢混合骨骼肌中分离出线粒体,这三种脊椎动物分别是实验大鼠褐家鼠、鬃狮蜥鬃狮蜥和甘蔗蟾蜍海蟾蜍。这些脊椎动物的体重大致相同,体温也相似。通过氧化还原差异光谱法测量这些分离出线粒体中细胞色素B、C、C1和A的含量。用羧基苍术苷滴定线粒体呼吸来测量腺嘌呤核苷酸转位酶(ANT),同时还测量分离出线粒体的蛋白质和磷脂含量。测量线粒体磷脂的脂肪酸组成。在37摄氏度下,在状态III和IV条件下以及在寡霉素抑制期间测量线粒体呼吸。不同物种的线粒体细胞色素比例不同。肌肉线粒体与肾脏和肝脏线粒体的不同之处在于,相对于细胞色素含量,其ANT含量更高。将呼吸速率相对于多个分母进行比较,发现相对于线粒体蛋白质含量表示时变化最大,而相对于线粒体细胞色素A和ANT含量表示时变化最小。计算细胞色素的周转,发现其在1至94电子秒(-1)之间变化。发现线粒体细胞色素的分子活性与线粒体膜脂质的相对多不饱和程度显著正相关。