Identification of the protein responsible for pyruvate transport into rat liver and heart mitochondria by specific labelling with [3H]N-phenylmaleimide.
نویسندگان
چکیده
1. N-Phenylmaleimide irreversibly inhibits pyruvate transport into rat heart and liver mitochondria to a much greater extent than does N-ethylmaleimide, iodoacetate or bromopyruvate. alpha-Cyanocinnamate protects the pyruvate transporter from attack by this thiol-blocking reagent. 2. In both heart and liver mitochondria alpha-cyanocinnamate diminishes labelling by [3H]N-phenylmaleimide of a membrane protein of subunit mol.wt. 15000 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 3. Exposure of mitochondrial to unlabelled N-phenylmaleimide in the presence of alpha-cyanocinnamate, followed by removal of alpha-cyanocinnamate and exposure to [3H]N-phenylmaleimide, produced specific labelling of the same protein. 4. Both labelling and kinetic experiments with inhibitors gave values for the approximate amount of carrier present in liver and heart mitochondria of 100 and 450 pmol/mg of mitochondrial protein respectively. 5. The turnover numbers for net pyruvate transport and pyruvate exchange at 0 degrees C were 6 and 200 min-1 respectively.
منابع مشابه
Identification of Intracellular Sources Responsible for Endogenous Reactive Oxygen Species Formation
The endogenous reactive oxygen species ("ROS") formation is associated with many pathologic states such as inflammatory diseases, neurodegenerative diseases, brain and heart ischemic injuries, cancer, and aging. The purpose of this study was to investigate the endogenous sources for "ROS" formation in intact isolated rat hepatocytes, in particular, peroxisomal oxidases, monoamine oxidase, xanth...
متن کاملIdentification of Intracellular Sources Responsible for Endogenous Reactive Oxygen Species Formation
The endogenous reactive oxygen species ("ROS") formation is associated with many pathologic states such as inflammatory diseases, neurodegenerative diseases, brain and heart ischemic injuries, cancer, and aging. The purpose of this study was to investigate the endogenous sources for "ROS" formation in intact isolated rat hepatocytes, in particular, peroxisomal oxidases, monoamine oxidase, xanth...
متن کاملThe effect of 6-deoxyclitoriacetal from Clitoria macrophylla Wall. on rat liver mitochondrial respiration and ATPase activity
A Thai plant Clitoria macrophylla Wall. was claimed to possess some pharmacological activities. The phytochemical studies reported a rotenoid compound, 6-deoxyclitoriacetal, which exhibits the cytotoxic effect in several cell line experiments. This report was to study the effect of 6-deoxyclitoriacetal on rat liver mitochondria, which may associated to its cytotoxic phenomena. Mitochondrial sus...
متن کاملThe effect of 6-deoxyclitoriacetal from Clitoria macrophylla Wall. on rat liver mitochondrial respiration and ATPase activity
A Thai plant Clitoria macrophylla Wall. was claimed to possess some pharmacological activities. The phytochemical studies reported a rotenoid compound, 6-deoxyclitoriacetal, which exhibits the cytotoxic effect in several cell line experiments. This report was to study the effect of 6-deoxyclitoriacetal on rat liver mitochondria, which may associated to its cytotoxic phenomena. Mitochondrial sus...
متن کاملThe concentration of the mitochondrial pyruvate carrier in rat liver and heart mitochondria determined with xc-cyano-#-(l-phenylindol-3-yl)acrylate
1. a-Cyano-f-(1-phenylindol-3-yl)acrylate inhibited pyruvate transport into both liver and heart mitochondria approximately linearly with respect to its concentration until 65% inhibition was achieved. 2. The extent of inhibition was dependent on the mitochondrial protein concentration. 3. By extrapolation of plots of inhibition versus inhibitor concentration to total inhibition, or by mathemat...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- The Biochemical journal
دوره 196 2 شماره
صفحات -
تاریخ انتشار 1981