Soares Nelson C, Spät Philipp, Méndez Jose Antonio, Nakedi Kehilwe, Aranda Jesús, Bou German
Proteome Center Tuebingen, University of Tuebingen, Germany; Laboratorio de Microbiología, Instituto de Investigación Biomedica de A Coruña (INIBIC), Servicio de Microbiología, Complejo Hospitalario Universitario A Coruña (CHUAC), Spain; Division of Medical Biochemistry (Dep. of Clinical Laboratory Sciences), Institute of Infectious Disease and Molecular Medicine, University of Cape Town-Medical School, Anzio, Observatory, Cape Town 7925, South Africa.
Proteome Center Tuebingen, University of Tuebingen, Germany.
J Proteomics. 2014 May 6;102:113-24. doi: 10.1016/j.jprot.2014.03.009. Epub 2014 Mar 21.
In the current study, the Ser/Thr/Tyr phosphoproteomes of two Acinetobacter baumannii strains, reference (ATCC17978) and highly invasive multidrug-resistant clinical isolate (Abh12O-A2) were analyzed using SCX and TiO2 chromatography followed by high resolution mass spectrometry. We detected a total of 201 unique phosphorylation sites (p-sites), and, after manual validation of peptide spectra, 91 high-confidence phosphorylation events (p-events) could be localized to a specific amino acid residue. The percentage distribution of Ser/Thr/Tyr phosphorylation was 68.9% on serine, 24.1% on threonine and 5.2% on tyrosine in ATCC17978, and 70.8% on serine, 25.2% on threonine and 3.8% on tyrosine in AbH12O-A2. Across all identified p-sites, 11 were identified in ATCC17978 only, while 43 were identified in Abh12O-A2 only, and 37 overlapped between the two strains. Here for the first time we describe the phosphoproteome of A. baumanii, and significance of selected phosphorylation sites is discussed in the context of stress/starvation, pathogenicity and drug resistance.
It is now well established that protein phosphorylation on Ser/Thr/Tyr residues is an important post-translational modification in bacteria. Herein we employed SCX and TiO2 chromatographic phosphopeptide enrichment combined with LTQ-Orbitrap mass spectrometric analyses to characterize and establish a qualitative comparison between the Ser/Thr/Tyr phosphoproteomes of two Acinetobacter baumannii strains: a reference strain and a highly invasive multidrug-resistant clinical isolate. We highlight the identification of phosphoproteins with a role in pathogenicity and those involved in drug resistance.
在本研究中，使用强阳离子交换色谱（SCX）和二氧化钛（TiO₂）色谱，随后进行高分辨率质谱分析，对两株鲍曼不动杆菌（参考菌株ATCC17978和高侵袭性多重耐药临床分离株Abh12O - A2）的丝氨酸/苏氨酸/酪氨酸磷酸化蛋白质组进行了分析。我们总共检测到201个独特的磷酸化位点（p位点），并且在对肽谱进行人工验证后，91个高可信度的磷酸化事件（p事件）可定位到特定的氨基酸残基。在ATCC17978中，丝氨酸、苏氨酸和酪氨酸磷酸化的百分比分布分别为68.9%、24.1%和5.2%；在AbH12O - A2中，丝氨酸、苏氨酸和酪氨酸磷酸化的百分比分布分别为7O.8%、25.2%和3.8%。在所有鉴定出的p位点中，11个仅在ATCC17978中鉴定到，43个仅在Abh12O - A2中鉴定到，37个在两株菌株中重叠。在此我们首次描述了鲍曼不动杆菌的磷酸化蛋白质组，并在应激/饥饿、致病性和耐药性的背景下讨论了所选磷酸化位点的意义。
目前已经明确，丝氨酸/苏氨酸/酪氨酸残基上的蛋白质磷酸化是细菌中一种重要的翻译后修饰。在此，我们采用SCX和TiO₂色谱磷酸肽富集方法，结合线性离子阱-轨道阱质谱分析，对两株鲍曼不动杆菌（一株参考菌株和一株高侵袭性多重耐药临床分离株）的丝氨酸/苏氨酸/酪氨酸磷酸化蛋白质组进行表征并建立定性比较。我们着重鉴定了在致病性和耐药性中起作用的磷酸化蛋白质。
