Hundal H S, Marette A, Ramlal T, Liu Z, Klip A
Division of Cell Biology, Hospital for Sick Children, Toronto, Ont. Canada.
FEBS Lett. 1993 Aug 16;328(3):253-8. doi: 10.1016/0014-5793(93)80938-q.
Hindlimb skeletal muscles of the rat express two isoforms of the alpha (alpha 1 and alpha 2) and two isoforms of the beta (beta 1 and beta 2) subunits of the Na+,K(+)-ATPase. Because several muscles constitute the hindlimb, we investigated if specific isoforms are expressed in particular muscles. Northern blot analysis using isoform-specific cDNA probes demonstrated that soleus muscle expressed only the beta 1 transcript, whereas EDL or white gastrocnemius muscles expressed only the beta 2 transcript, and red gastrocnemius muscle expressed both mRNAs. All muscles tested expressed both alpha 1 and alpha 2 transcripts, albeit to various degrees: alpha 1 transcripts were present to about the same extent in all muscles but alpha 2 mRNA was 4-fold more abundant in soleus than in EDL for the same amount of total RNA. Beta subunit protein levels were investigated in purified plasma membrane fractions of pooled red (soleus + red gastrocnemius + red quadriceps) or white (white gastrocnemius + white quadriceps) muscles using isoform-specific antibodies. Red muscles expressed mostly the beta 1 protein while white muscles expressed mostly the beta 2 subunit. Both muscle groups had similar levels of alpha 1 or alpha 2 subunits, and crude membranes isolated from red muscles had 30% higher Na+,K(+)-ATPase activity than white muscle membranes. We conclude that oxidative muscles (slow and fast twitch) express beta 1 subunits, whereas glycolytic, fast twitch muscles express beta 2 subunits, and that both beta isoforms support the Na+,K(+)-ATPase activity of the alpha subunits.
大鼠后肢骨骼肌表达Na⁺,K⁺-ATP酶α亚基的两种同工型(α1和α2)以及β亚基的两种同工型(β1和β2)。由于后肢由几块肌肉组成,我们研究了特定的同工型是否在特定肌肉中表达。使用同工型特异性cDNA探针进行的Northern印迹分析表明,比目鱼肌仅表达β1转录本,而趾长伸肌或白色腓肠肌仅表达β2转录本,红色腓肠肌则同时表达这两种mRNA。所有测试的肌肉均表达α1和α2转录本,尽管程度不同:在相同总量的总RNA中,α1转录本在所有肌肉中的含量大致相同,但α2 mRNA在比目鱼肌中的丰度比在趾长伸肌中高4倍。使用同工型特异性抗体,在合并的红色(比目鱼肌+红色腓肠肌+红色股四头肌)或白色(白色腓肠肌+白色股四头肌)肌肉的纯化质膜组分中研究了β亚基蛋白水平。红色肌肉主要表达β1蛋白,而白色肌肉主要表达β2亚基。两组肌肉的α1或α2亚基水平相似,并且从红色肌肉分离的粗膜的Na⁺,K⁺-ATP酶活性比白色肌肉膜高30%。我们得出结论,氧化型肌肉(慢肌和快肌)表达β1亚基,而糖酵解型快肌表达β2亚基,并且两种β同工型均支持α亚基的Na⁺,K⁺-ATP酶活性。