Plant peptidoglycan precursor biosynthesis: Conservation between moss chloroplasts and Gram-negative bacteria.
Plant peptidoglycan precursor biosynthesis: Conservation between moss chloroplasts and Gram-negative bacteria.
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DOI：
10.1093/plphys/kiac176
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发表时间：
2022-08-29
期刊：
影响因子：
7.4
通讯作者：
Dowson, Christopher G.
中科院分区：
文献类型：
--
作者：
Dowson, Amanda J.;Lloyd, Adrian J.;Cuming, Andrew C.;Roper, David, I;Frigerio, Lorenzo;Dowson, Christopher G.
越来越多的证据表明，与细菌细胞壁一致的肽聚糖在许多光合真核生物的叶绿体周围合成，从蓝绿藻到早期分化的陆地植物，包括蕨类植物，但生物合成途径尚未得到证实。在这里，我们采用了质谱和酶学在一个双重的方法来表征的肽聚糖在叶绿体的藓类立碗藓（立碗藓）patens的合成。为了驱动肽聚糖途径中间体的积累，将展布假单胞菌与抗生素磷霉素、D-环丝氨酸和羧苄青霉素一起培养，这些抗生素抑制细菌中的关键肽聚糖途径蛋白。三氯乙酸提取的苔藓代谢组的质谱分析显示，从尿苷二磷酸N-乙酰葡糖胺（UDP-GlcNAc）到尿苷二磷酸N-乙酰胞壁酸（UDP-MurNAc）-D，L-二氨基庚二酸（DAP）-五肽的五种预测中间体水平升高。大多数革兰氏阴性细菌，包括蓝细菌，将内消旋二氨基庚二酸（D，L-DAP）掺入肽聚糖的茎肽的第三个残基中，而不是大多数革兰氏阳性细菌典型的L-赖氨酸。为了确定D，L-DAP掺入到P. patens前体中的特异性，我们分析了来自P. patens和蓝细菌鱼腥藻（Nostoc sp.菌株PCC 7120）的重组蛋白UDP-N-乙酰胞壁酰-L-丙氨酰-D-谷氨酸-2，6-二氨基庚二酸连接酶（MurE）。这两种连接酶掺入D，L-DAP几乎完全优先于L-Lys，与质谱数据一致，催化效率类似于以前记录的革兰氏阴性细菌MurE连接酶。我们讨论了这些数据如何符合雅阁与DL-DAP结合细菌MurE连接酶和植物肽聚糖途径内的水平基因转移事件的概率的活性位点残基的保护。早分化陆地植物中的功能性肽聚糖生物合成途径源自叶绿体的革兰氏阴性细菌祖细胞。
Accumulating evidence suggests that peptidoglycan, consistent with a bacterial cell wall, is synthesized around the chloroplasts of many photosynthetic eukaryotes, from glaucophyte algae to early-diverging land plants including pteridophyte ferns, but the biosynthetic pathway has not been demonstrated. Here, we employed mass spectrometry and enzymology in a two-fold approach to characterize the synthesis of peptidoglycan in chloroplasts of the moss Physcomitrium (Physcomitrella) patens. To drive the accumulation of peptidoglycan pathway intermediates, P. patens was cultured with the antibiotics fosfomycin, D-cycloserine, and carbenicillin, which inhibit key peptidoglycan pathway proteins in bacteria. Mass spectrometry of the trichloroacetic acid-extracted moss metabolome revealed elevated levels of five of the predicted intermediates from uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) through the uridine diphosphate N-acetylmuramic acid (UDP-MurNAc)-D,L-diaminopimelate (DAP)-pentapeptide. Most Gram-negative bacteria, including cyanobacteria, incorporate meso-diaminopimelic acid (D,L-DAP) into the third residue of the stem peptide of peptidoglycan, as opposed to L-lysine, typical of most Gram-positive bacteria. To establish the specificity of D,L-DAP incorporation into the P. patens precursors, we analyzed the recombinant protein UDP-N-acetylmuramoyl-L-alanyl-D-glutamate–2,6-diaminopimelate ligase (MurE) from both P. patens and the cyanobacterium Anabaena sp. (Nostoc sp. strain PCC 7120). Both ligases incorporated D,L-DAP in almost complete preference to L-Lys, consistent with the mass spectrophotometric data, with catalytic efficiencies similar to previously documented Gram-negative bacterial MurE ligases. We discuss how these data accord with the conservation of active site residues common to DL-DAP-incorporating bacterial MurE ligases and of the probability of a horizontal gene transfer event within the plant peptidoglycan pathway. A functional peptidoglycan biosynthetic pathway in early-diverging land plants is derived from a Gram-negative bacterial progenitor of chloroplasts.
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DOI：
10.1046/j.0014-2956.2001.02524.x
发表时间：
2001-11-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Dementin, S;Bouhss, A;Blanot, D
通讯作者：
Blanot, D
影响因子：
5.7
作者：
De Benedetti, Stefania;Buehl, Henrike;Henrichfreise, Beate
通讯作者：
Henrichfreise, Beate
DOI：
10.1073/pnas.0510693103
发表时间：
2006-04-25
影响因子：
11.1
作者：
Machida, M;Takechi, K;Takano, H
通讯作者：
Takano, H
影响因子：
7.2
作者：
Lang, Daniel;Ullrich, Kristian K.;Rensing, Stefan A.
通讯作者：
Rensing, Stefan A.
影响因子：
4.8
作者：
Boniface, Audrey;Bouhss, Ahmed;Blanot, Didier
通讯作者：
Blanot, Didier