title: Proteome profile of functional mitochondria from human skeletal muscle using one-dimensional gel electrophoresis and HPLC-ESI-MS/MS.
abstract: Mitochondria can be isolated from skeletal muscle in a manner that preserves tightly coupled bioenergetic function in vitro. The purpose of this study was to characterize the composition of such preparations using a proteomics approach. Mitochondria isolated from human vastus lateralis biopsies were functional as evidenced by their response to carbohydrate and fat-derived fuels. Using one-dimensional gel electrophoresis and HPLC-ESI-MS/MS, 823 unique proteins were detected, and 487 of these were assigned to the mitochondrion, including the newly characterized SIRT5, MitoNEET and RDH13. Proteins detected included 9 of the 13 mitochondrial DNA-encoded proteins and 86 of 104 electron transport chain (ETC) and ETC-related proteins. In addition, 59 of 78 proteins of the 55S mitoribosome, several TIM and TOM proteins and cell death proteins were present. This study presents an efficient method for future qualitative assessments of proteins from functional isolated mitochondria from small samples of healthy and diseased skeletal muscle.
authors: Center for Metabolic Biology, Arizona State University, Tempe, AZ, USA.
authors: Natalie
authors: N
authors: Lefort
authors: Zhengping
authors: Z
authors: Yi
authors: Benjamin
authors: B
authors: Bowen
authors: Brian
authors: B
authors: Glancy
authors: Eleanna A
authors: EA
authors: De Filippis
authors: Rebekka
authors: R
authors: Mapes
authors: Hyonson
authors: H
authors: Hwang
authors: Charles R
authors: CR
authors: Flynn
authors: Wayne T
authors: WT
authors: Willis
authors: Anthony
authors: A
authors: Civitarese
authors: Kurt
authors: K
authors: Højlund
authors: Lawrence J
authors: LJ
authors: Mandarino
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004579:Electron Transport; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005227:Fatty Acids / Q000737:chemistry; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D018384:Oxidative Stress; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000596:Amino Acids; D005227:Fatty Acids; D011506:Proteins