TY - JOUR
T1 - Proteome profile of functional mitochondria from human skeletal muscle using one-dimensional gel electrophoresis and HPLC-ESI-MS/MS
AU - Lefort, Natalie
AU - Yi, Zhengping
AU - Bowen, Benjamin
AU - Glancy, Brian
AU - De Filippis, Eleanna A.
AU - Mapes, Rebekka
AU - Hwang, Hyonson
AU - Flynn, Charles R.
AU - Willis, Wayne T.
AU - Civitarese, Anthony
AU - Højlund, Kurt
AU - Mandarino, Lawrence J.
PY - 2009/8/20
Y1 - 2009/8/20
N2 - 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.
AB - 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.
KW - HPLC-ESI-MS/MS
KW - Human skeletal muscle
KW - Mitochondria
KW - One-dimensional-gel electrophoresis
KW - Proteome
UR - http://www.scopus.com/inward/record.url?scp=67651151024&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67651151024&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2009.06.011
DO - 10.1016/j.jprot.2009.06.011
M3 - Article
C2 - 19567276
AN - SCOPUS:67651151024
SN - 1874-3919
VL - 72
SP - 1046
EP - 1060
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 6
ER -