Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles.
Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles.
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DOI：
10.1093/glycob/cwy100
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发表时间：
2019-01-01
期刊：
影响因子：
4.3
通讯作者：
Juge N
中科院分区：
文献类型：
--
作者：
Latousakis D;Nepravishta R;Rejzek M;Wegmann U;Le Gall G;Kavanaugh D;Colquhoun IJ;Frese S;MacKenzie DA;Walter J;Angulo J;Field RA;Juge N
罗伊氏乳杆菌是一种寄生在许多脊椎动物胃肠道中的肠道共生体。表面暴露的富丝氨酸重复蛋白（SRRP）是革兰氏阳性菌的主要黏附蛋白。通过对野生型或罗伊氏乳杆菌等基因突变体的凝集素和糖核苷酸谱分析，以及对srrp的MALDI-ToF-MS、LC-MS和GC-MS分析，我们发现罗伊氏乳杆菌菌株100-23C（来自啮齿动物）和ATCC 53608（来自猪）可以进行蛋白o糖基化，并分别用hexx - glc - glcnac和di-GlcNAc修饰SRRP100-23和SRRP53608。此外，大肠杆菌体内糖工程导致SRRP53608变体α-GlcNAc和GlcNAcβ(1→6)GlcNAcα片段糖基化。在SecA2/Y2辅助分泌系统中鉴定了参与这些粘附素修饰的糖基转移酶，并通过饱和转移差核磁共振光谱和差示扫描荧光法确定了它们的糖核苷酸偏好。总之，这些发现为肠道共生菌的细胞o蛋白糖基化途径和糖工程应用的潜在途径提供了新的见解。
Lactobacillus reuteri is a gut symbiont inhabiting the gastrointestinal tract of numerous vertebrates. The surface-exposed serine-rich repeat protein (SRRP) is a major adhesin in Gram-positive bacteria. Using lectin and sugar nucleotide profiling of wild-type or L. reuteri isogenic mutants, MALDI-ToF-MS, LC–MS and GC–MS analyses of SRRPs, we showed that L. reuteri strains 100-23C (from rodent) and ATCC 53608 (from pig) can perform protein O-glycosylation and modify SRRP100-23 and SRRP53608 with Hex-Glc-GlcNAc and di-GlcNAc moieties, respectively. Furthermore, in vivo glycoengineering in E. coli led to glycosylation of SRRP53608 variants with α-GlcNAc and GlcNAcβ(1→6)GlcNAcα moieties. The glycosyltransferases involved in the modification of these adhesins were identified within the SecA2/Y2 accessory secretion system and their sugar nucleotide preference determined by saturation transfer difference NMR spectroscopy and differential scanning fluorimetry. Together, these findings provide novel insights into the cellular O-protein glycosylation pathways of gut commensal bacteria and potential routes for glycoengineering applications.
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影响因子：
5.6
作者：
Latousakis D;Juge N
通讯作者：
Juge N
影响因子：
4.5
作者：
Frese SA;Mackenzie DA;Peterson DA;Schmaltz R;Fangman T;Zhou Y;Zhang C;Benson AK;Cody LA;Mulholland F;Juge N;Walter J
通讯作者：
Walter J
影响因子：
5.1
作者：
Etzold, Sabrina;Kober, Olivia I.;Juge, Nathalie
通讯作者：
Juge, Nathalie
影响因子：
7
作者：
Chaze, Thibault;Guillot, Alain;Mistou, Michel-Yves
通讯作者：
Mistou, Michel-Yves
影响因子：
3.6
作者：
Bensing, BA;Sullam, PM
通讯作者：
Sullam, PM