BACKGROUND: The measurement of the activities of the five complexes comprising the respiratory chain has proven to be a major challenge when a limiting amount of biological material is available. Here we report a set of three convenient assays that allows this measurement under such circumstances. METHODS: One assay relies on the sequential addition of reagents to measure first complex IV activity, followed by complex II+III, and then glycerol-3-phosphate dehydrogenase+complex III activities and finally isolated complex III activity. A second assay measures the activity of complex II followed by glycerol-3-phosphate dehydrogenase and isocitrate dehydrogenase. A third assay measures rotenone-sensitive complex I activity and subsequently oligomycin-sensitive complex V activity. RESULTS: These assays have been successfully used on extracts of small numbers of human cells displaying various defects in the respiratory chain, and on frozen tissue homogenates of retina and very early mouse embryos. CONCLUSIONS: The strength of this set of assays lies both in its rapid and simple execution and its capacity for immediate detection of partial defects, because each activity can be compared with one or two other activities measured in the same sample.
BACKGROUND: The measurement of the activities of the five complexes comprising the respiratory chain has proven to be a major challenge when a limiting amount of biological material is available. Here we report a set of three convenient assays that allows this measurement under such circumstances. METHODS: One assay relies on the sequential addition of reagents to measure first complex IV activity, followed by complex II+III, and then glycerol-3-phosphate dehydrogenase+complex III activities and finally isolated complex III activity. A second assay measures the activity of complex II followed by glycerol-3-phosphate dehydrogenase and isocitrate dehydrogenase. A third assay measures rotenone-sensitive complex I activity and subsequently oligomycin-sensitive complex V activity. RESULTS: These assays have been successfully used on extracts of small numbers of human cells displaying various defects in the respiratory chain, and on frozen tissue homogenates of retina and very early mouse embryos. CONCLUSIONS: The strength of this set of assays lies both in its rapid and simple execution and its capacity for immediate detection of partial defects, because each activity can be compared with one or two other activities measured in the same sample.
Authors: Huiliang Zhang; Pei Wang; Sara Bisetto; Yisang Yoon; Quan Chen; Shey-Shing Sheu; Wang Wang Journal: Cardiovasc Res Date: 2016-10-29 Impact factor: 10.787
Authors: Katherine A Janeway; Su Young Kim; Maya Lodish; Vânia Nosé; Pierre Rustin; José Gaal; Patricia L M Dahia; Bernadette Liegl; Evan R Ball; Margarita Raygada; Angela H Lai; Lorna Kelly; Jason L Hornick; Maureen O'Sullivan; Ronald R de Krijger; Winand N M Dinjens; George D Demetri; Cristina R Antonescu; Jonathan A Fletcher; Lee Helman; Constantine A Stratakis Journal: Proc Natl Acad Sci U S A Date: 2010-12-20 Impact factor: 11.205
Authors: Glenn C Rowe; Ian S Patten; Zsuzsanna K Zsengeller; Riyad El-Khoury; Mitsuharu Okutsu; Sophia Bampoh; Nicole Koulisis; Caitlin Farrell; Michael F Hirshman; Zhen Yan; Laurie J Goodyear; Pierre Rustin; Zolt Arany Journal: Cell Rep Date: 2013-05-23 Impact factor: 9.423
Authors: Led Yered Jafet García Montes de Oca; Tecilli Cabellos Avelar; Gerardo Ignacio Picón Garrido; Alicia Chagoya-López; Luis González de la Vara; Norma Laura Delgado Buenrostro; Yolanda Irasema Chirino-López; Carlos Gómez-Lojero; Emma Berta Gutiérrez-Cirlos Journal: J Bioenerg Biomembr Date: 2016-08-09 Impact factor: 2.945