Penyige András, Keseru Judit, Fazakas Ferenc, Schmelczer Iván, Szirák Krisztina, Barabás György, Biró Sándor
Department of Human Genetics, University of Debrecen, Debrecen, Hungary.
J Microbiol. 2009 Oct;47(5):549-56. doi: 10.1007/s12275-009-0032-y. Epub 2009 Oct 24.
Mono-ADP-ribosylation is the enzymatic transfer of ADP-ribose from NAD(+) to acceptor proteins catalyzed by ADP-ribosyltransferases. Using m-aminophenylboronate affinity chromatography, 2D-gel electrophoresis, in-gel digestion and MALDI-TOF analysis we have identified eight in vitro ADP-ribosylated proteins in Streptomyces coelicolor, which can be classified into three categories: (i) secreted proteins; (ii) metabolic enzymes using NAD(+)/NADH or NADP(+)/NADPH as coenzymes; and (iii) other proteins. The secreted proteins could be classified into two functional categories: SCO2008 and SC05477 encode members of the family of periplasmic extracellular solute-binding proteins, and SCO6108 and SC01968 are secreted hydrolases. Dehydrogenases are encoded by SC04824 and SC04771. The other targets are GlnA (glutamine synthetase I., SC02198) and SpaA (starvation-sensing protein encoded by SC07629). SCO2008 protein and GlnA had been identified as ADP-ribosylated proteins in previous studies. With these results we provided experimental support for a previous suggestion that ADP-ribosylation may regulate membrane transport and localization of periplasmic proteins. Since ADP-ribosylation results in inactivation of the target protein, ADP-ribosylation of dehydrogenases might modulate crucial primary metabolic pathways in Streptomyces. Several of the proteins identified here could provide a strong connection between protein ADP-ribosylation and the regulation of morphological differentiation in S. coelicolor.
单（ADP - 核糖）基化是由ADP - 核糖基转移酶催化将ADP - 核糖从NAD⁺转移至受体蛋白的酶促反应。我们利用间氨基苯硼酸亲和色谱、二维凝胶电泳、胶内酶解和基质辅助激光解吸电离飞行时间质谱分析，在天蓝色链霉菌中鉴定出8种体外ADP - 核糖基化蛋白，这些蛋白可分为三类：（i）分泌蛋白；（ii）以NAD⁺/NADH或NADP⁺/NADPH为辅酶的代谢酶；（iii）其他蛋白。分泌蛋白可分为两个功能类别：SCO2008和SC05477编码周质细胞外溶质结合蛋白家族成员，SCO6108和SC01968是分泌型水解酶。脱氢酶由SC04824和SC04771编码。其他靶点是GlnA（谷氨酰胺合成酶I，SC02198）和SpaA（由SC07629编码的饥饿感应蛋白）。SCO2008蛋白和GlnA在先前的研究中已被鉴定为ADP - 核糖基化蛋白。基于这些结果，我们为先前的一个观点提供了实验支持，即ADP - 核糖基化可能调节周质蛋白的膜转运和定位。由于ADP - 核糖基化导致靶蛋白失活，脱氢酶的ADP - 核糖基化可能会调节链霉菌中关键的初级代谢途径。这里鉴定出的几种蛋白可能在蛋白ADP - 核糖基化与天蓝色链霉菌形态分化调控之间建立紧密联系。
