Börngen Kirsten, Battle Andrew R, Möker Nina, Morbach Susanne, Marin Kay, Martinac Boris, Krämer Reinhard
Institute of Biochemistry, University of Cologne, 50674 Cologne, Germany.
Biochim Biophys Acta. 2010 Nov;1798(11):2141-9. doi: 10.1016/j.bbamem.2010.06.022. Epub 2010 Jul 1.
Based on sequence similarity, the mscCG gene product of Corynebacterium glutamicum belongs to the family of MscS-type mechanosensitive channels. In order to investigate the physiological significance of MscCG in response to osmotic shifts in detail, we studied its properties using both patch-clamp techniques and betaine efflux kinetics. After heterologous expression in an Escherichiacoli strain devoid of mechanosensitive channels, in patch-clamp analysis of giant E. coli spheroplasts MscCG showed the typical pressure dependent gating behavior of a stretch-activated channel with a current/voltage dependence indicating a strongly rectifying behavior. Apart from that, MscCG is characterized by significant functional differences with respect to conductance, ion selectivity and desensitation behavior as compared to MscS from E. coli. Deletion and complementation studies in C. glutamicum showed a significant contribution of MscCG to betaine efflux in response to hypoosmotic conditions. A detailed analysis of concomitant betaine uptake (by the betaine transporter BetP) and efflux (by MscCG) under hyperosmotic conditions indicates that MscCG may act in osmoregulation in C. glutamicum by fine-tuning the steady state concentration of compatible solutes in the cytoplasm which are accumulated in response to hyperosmotic stress.
基于序列相似性,谷氨酸棒杆菌的mscCG基因产物属于MscS型机械敏感通道家族。为了详细研究MscCG在应对渗透压变化时的生理意义,我们使用膜片钳技术和甜菜碱外排动力学研究了它的特性。在缺乏机械敏感通道的大肠杆菌菌株中进行异源表达后,在对巨大的大肠杆菌原生质球进行膜片钳分析时,MscCG表现出拉伸激活通道典型的压力依赖性门控行为,其电流/电压依赖性表明具有强整流行为。除此之外,与大肠杆菌的MscS相比,MscCG在电导、离子选择性和脱敏行为方面具有显著的功能差异。谷氨酸棒杆菌中的缺失和互补研究表明,MscCG在低渗条件下对甜菜碱外排有显著贡献。对高渗条件下伴随的甜菜碱摄取(通过甜菜碱转运体BetP)和外排(通过MscCG)的详细分析表明,MscCG可能通过微调细胞质中相容性溶质的稳态浓度来参与谷氨酸棒杆菌的渗透调节,这些溶质是在高渗应激下积累的。