Department of Biochemistry and Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
Department of Botany, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-783 71 Olomouc, Czech Republic.
Molecules. 2021 Mar 17;26(6):1683. doi: 10.3390/molecules26061683.
The aim and novelty of this paper are found in assessing the influence of inhibitors and antibiotics on intact cell MALDI-TOF mass spectra of the cyanobacterium sp. UPOC S4 and to check the impact on reliability of identification. Defining the limits of this method is important for its use in biology and applied science. The compounds included inhibitors of respiration, glycolysis, citrate cycle, and proteosynthesis. They were used at 1-10 μM concentrations and different periods of up to 3 weeks. Cells were also grown without inhibitors in a microgravity because of expected strong effects. Mass spectra were evaluated using controls and interpreted in terms of differential peaks and their assignment to protein sequences by mass. Antibiotics, azide, and bromopyruvate had the greatest impact. The spectral patterns were markedly altered after a prolonged incubation at higher concentrations, which precluded identification in the database of reference spectra. The incubation in microgravity showed a similar effect. These differences were evident in dendrograms constructed from the spectral data. Enzyme inhibitors affected the spectra to a smaller extent. This study shows that only a long-term presence of antibiotics and strong metabolic inhibitors in the medium at 10 M concentrations hinders the correct identification of cyanobacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF).
本文的目的和新颖性在于评估抑制剂和抗生素对蓝藻 sp. UPOC S4 完整细胞 MALDI-TOF 质谱的影响,并检查其对鉴定可靠性的影响。定义该方法的局限性对于其在生物学和应用科学中的应用非常重要。所包括的化合物有呼吸、糖酵解、柠檬酸循环和蛋白质合成抑制剂。它们以 1-10 μM 的浓度和长达 3 周的不同时间使用。由于预期会产生强烈影响,细胞也在微重力下生长而没有抑制剂。使用对照评估质谱,并根据差异峰及其通过质量分配到蛋白质序列来解释。抗生素、叠氮化钠和溴丙酮酸的影响最大。在较高浓度下长时间孵育后,光谱模式发生明显改变,这使得在参考光谱数据库中无法进行鉴定。在微重力下孵育也显示出类似的效果。这些差异在从光谱数据构建的聚类图中显而易见。酶抑制剂对光谱的影响较小。这项研究表明,只有在培养基中存在长期的抗生素和强代谢抑制剂,浓度为 10 μM,才会阻碍基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF)对蓝藻的正确鉴定。