| Autor: |
Smith CD; Department of Physiology and Developmental Biology, Brigham Young University, Provo, Utah, USA., Compton RA, Bowler JS, Kemp JT, Sudweeks SN, Thomson DM, Winder WW |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of applied physiology (Bethesda, Md. : 1985) [J Appl Physiol (1985)] 2011 Dec; Vol. 111 (6), pp. 1622-8. Date of Electronic Publication: 2011 Sep 08. |
| DOI: |
10.1152/japplphysiol.00160.2011 |
| Abstrakt: |
In liver, the AMP-activated protein kinase kinase (AMPKK) complex was identified as the association of liver kinase B1 (LKB1), mouse protein 25 (MO25α/β), and Ste-20-related adaptor protein (STRADα/β); however, this complex has yet to be characterized in skeletal muscle. We demonstrate the expression of the LKB1-MO25-STRAD complex in skeletal muscle, confirm the absence of mRNA splice variants, and report the relative mRNA expression levels of these proteins in control and muscle-specific LKB1 knockout (LKB1(-/-)) mouse muscle. LKB1 detection in untreated control and LKB1(-/-) muscle lysates revealed two protein bands (50 and 60 kDa), although only the heavier band was diminished in LKB1(-/-) samples [55 ± 2.5 and 13 ± 1.5 arbitrary units (AU) in control and LKB1(-/-), respectively, P < 0.01], suggesting that LKB1 is not represented at 50 kDa, as previously cited. The 60-kDa LKB1 band was further confirmed following purification using polyethylene glycol (43 ± 5 and 8.4 ± 4 AU in control and LKB1(-/-), respectively, P < 0.01) and ion-exchange fast protein liquid chromatography. Mass spectrometry confirmed LKB1 protein detection in the 60-kDa protein band, while none was detected in the 50-kDa band. Coimmunoprecipitation assays demonstrated LKB1-MO25-STRAD complex formation. Quantitative PCR revealed significantly reduced LKB1, MO25α, and STRADβ mRNA in LKB1(-/-) muscle. These findings demonstrate that the LKB1-MO25-STRAD complex is the principal AMPKK in skeletal muscle. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
