Arena Simona, D'Ambrosio Chiara, Renzone Giovanni, Rullo Rosario, Ledda Luigi, Vitale Filomena, Maglione Giuseppe, Varcamonti Mario, Ferrara Lino, Scaloni Andrea
Proteomics & Mass Spectrometry Laboratory, I.S.P.A.A.M., National Research Council, via Argine 1085, 80147 Naples, Italy.
Proteomics. 2006 Jan;6(1):181-92. doi: 10.1002/pmic.200402109.
Streptococcus thermophilus is a Gram-positive bacterium belonging to the group of lactic acid bacteria, among which several genera play an essential role in manufacture of food products. Recently, a genomic consortium sequenced and annotated its entire genome, which has been demonstrated to contain 1900 coding sequences. In this study, we have revealed the expression products of almost 200 different genes using a proteomic strategy combining 2-DE plus MALDI-TOF PMF and differential 1-DE plus muLC-ESI-IT-MS/MS. Thus, a number of cellular pathways related to important physiological processes were described at the proteomic level. Almost 50 genes were related to multiple electrophoretic species, whose heterogeneity was mainly due to variability in pI values. A 2-DE reference map obtained for lactose-grown cells was compared with those obtained after heat, cold, acid, oxidative and starvation stresses. Protein up/down-regulation measurements demonstrated that adaptation to different environmental challenges may involve the contribution of unique as well as combined physiological mechanisms. Common regulatory sites in the promoter region of genes whose expression was induced after stress were identified. These results provide a better comprehension of biochemical processes related to stress resistance in S. thermophilus, allowing defining the molecular bases of adaptative responses or markers for the identification of strains with potential industrial applications.
嗜热链球菌是一种革兰氏阳性菌,属于乳酸菌属,其中几个属在食品生产中起着至关重要的作用。最近,一个基因组联盟对其全基因组进行了测序和注释,结果表明该基因组包含1900个编码序列。在本研究中,我们采用二维电泳(2-DE)结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF PMF)以及一维差异电泳(1-DE)结合微液相色谱-电喷雾离子阱质谱(muLC-ESI-IT-MS/MS)的蛋白质组学策略,揭示了近200个不同基因的表达产物。因此,在蛋白质组水平上描述了许多与重要生理过程相关的细胞途径。近50个基因与多种电泳条带相关,其异质性主要归因于等电点(pI)值的变化。将乳糖培养细胞获得的二维电泳参考图谱与热、冷、酸、氧化和饥饿应激后获得的图谱进行了比较。蛋白质上调/下调测量表明,适应不同的环境挑战可能涉及独特以及联合的生理机制。确定了应激后表达被诱导的基因启动子区域中的常见调控位点。这些结果有助于更好地理解嗜热链球菌中与抗逆性相关的生化过程,从而确定适应性反应的分子基础或用于鉴定具有潜在工业应用价值菌株的标志物。