Streptococcus mutans protein synthesis during mixed-species biofilm development by high-throughput quantitative proteomics.
Streptococcus mutans protein synthesis during mixed-species biofilm development by high-throughput quantitative proteomics.
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DOI：
10.1371/journal.pone.0045795
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Koo H
中科院分区：
文献类型：
--
作者：
Klein MI;Xiao J;Lu B;Delahunty CM;Yates JR 3rd;Koo H
牙齿表面形成的生物膜由缠结在细胞外基质中的混合微生物群组成。口腔生物膜不断暴露于环境变化中，影响微生物组成、基质形成和毒力表达。变形链球菌和蔗糖是与致龋生物膜演变相关的关键调节剂。在这项研究中，我们使用高通量定量蛋白质组学方法来研究S。在蔗糖诱导的混合物种生物膜发育过程中，变形杆菌产生促进其建立和最佳存活的相关蛋白质。S.在限制碳水化合物的条件下，在唾液包被的羟基磷灰石表面上最初形成单独的或与内氏放线菌和口腔链球菌混合的变形菌。然后引入蔗糖（1%，w/v）以引起环境变化，并诱导生物膜积累。多维蛋白质鉴定技术（MudPIT）检测到了高达60%的S.生物膜内的变异体。与胞外多糖基质组装，代谢和压力适应过程相关的特定蛋白质是非常丰富的生物膜从早期到后期的发展阶段，蔗糖引入后的过渡。结果表明，S.在混合物种生物膜群落中的变形杆菌增加了与葡聚糖合成和重塑（gtfBC，dexA）和葡聚糖结合（gbpB）相关的特定基因的表达（P<0.05）。此外，S.变形杆菌上调特定的适应机制以科普酸性环境（F1 F0-ATP酶系统、脂肪酸生物合成、支链氨基酸代谢）和分子伴侣（GroEL）。有趣的是，当S.变形杆菌形成混合物种生物膜（相对于单一物种生物膜），这表明在其它生物体存在下，基质组装、存活和生物膜维持方面存在根本差异。我们的数据提供了关于S.变形杆菌优化其代谢，并适应/生存在混合物种群落中，以响应动态变化的环境。这反映了与该细菌在复杂生物膜内表达毒力相关的复杂生理过程。
Biofilms formed on tooth surfaces are comprised of mixed microbiota enmeshed in an extracellular matrix. Oral biofilms are constantly exposed to environmental changes, which influence the microbial composition, matrix formation and expression of virulence. Streptococcus mutans and sucrose are key modulators associated with the evolution of virulent-cariogenic biofilms. In this study, we used a high-throughput quantitative proteomics approach to examine how S. mutans produces relevant proteins that facilitate its establishment and optimal survival during mixed-species biofilms development induced by sucrose. Biofilms of S. mutans, alone or mixed with Actinomyces naeslundii and Streptococcus oralis, were initially formed onto saliva-coated hydroxyapatite surface under carbohydrate-limiting condition. Sucrose (1%, w/v) was then introduced to cause environmental changes, and to induce biofilm accumulation. Multidimensional protein identification technology (MudPIT) approach detected up to 60% of proteins encoded by S. mutans within biofilms. Specific proteins associated with exopolysaccharide matrix assembly, metabolic and stress adaptation processes were highly abundant as the biofilm transit from earlier to later developmental stages following sucrose introduction. Our results indicate that S. mutans within a mixed-species biofilm community increases the expression of specific genes associated with glucan synthesis and remodeling (gtfBC, dexA) and glucan-binding (gbpB) during this transition (P<0.05). Furthermore, S. mutans up-regulates specific adaptation mechanisms to cope with acidic environments (F1F0-ATPase system, fatty acid biosynthesis, branched chain amino acids metabolism), and molecular chaperones (GroEL). Interestingly, the protein levels and gene expression are in general augmented when S. mutans form mixed-species biofilms (vs. single-species biofilms) demonstrating fundamental differences in the matrix assembly, survival and biofilm maintenance in the presence of other organisms. Our data provide insights about how S. mutans optimizes its metabolism and adapts/survives within the mixed-species community in response to a dynamically changing environment. This reflects the intricate physiological processes linked to expression of virulence by this bacterium within complex biofilms.