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Many fructosamine 3-kinase homologues in bacteria are ribulosamine/erythrulosamine 3-kinases potentially involved in protein deglycation.

机译:细菌中的许多果糖胺3激酶同源物是核糖胺/赤藓糖胺3激酶,可能与蛋白质脱糖有关。

摘要

The purpose of this work was to identify the function of bacterial homologues of fructosamine 3-kinase (FN3K), a mammalian enzyme responsible for the removal of fructosamines from proteins. FN3K homologues were identified in approximately 200 (i.e. approximately 27%) of the sequenced bacterial genomes. In 11 of these genomes, from phylogenetically distant bacteria, the FN3K homologue was immediately preceded by a low-molecular-weight protein-tyrosine-phosphatase (LMW-PTP) homologue, which is therefore probably functionally related to the FN3K homologue. Five bacterial FN3K homologues (from Escherichia coli, Enterococcus faecium, Lactobacillus plantarum, Staphylococcus aureus and Thermus thermophilus) were overexpressed in E. coli, purified and their kinetic properties investigated. Four were ribulosamine/erythrulosamine 3-kinases acting best on free lysine and cadaverine derivatives, but not on ribulosamines bound to the alpha amino group of amino acids. They also phosphorylated protein-bound ribulosamines or erythrulosamines, but not protein-bound fructosamines, therefore having properties similar to those of mammalian FN3K-related protein. The E. coli FN3K homologue (YniA) was inactive on all tested substrates. The LMW-PTP of T. thermophilus, which forms an operon with an FN3K homologue, and an LMW-PTP of S. aureus (PtpA) were overexpressed in E. coli, purified and shown to dephosphorylate not only protein tyrosine phosphates, but protein ribulosamine 5-phosphates as well as free ribuloselysine 5-phosphate and erythruloselysine 4-phosphate. These LMW-PTPs were devoid of ribulosamine 3-phosphatase activity. It is concluded that most bacterial FN3K homologues are ribulosamine/erythrulosamine 3-kinases. They may serve, in conjunction with a phosphatase, to deglycate products of glycation formed from ribose 5-phosphate or erythrose 4-phosphate.
机译:这项工作的目的是确定果糖胺3激酶(FN3K)的细菌同源物的功能,果糖胺3激酶是负责从蛋白质中去除果糖胺的哺乳动物酶。在约200个(即约27%)的测序细菌基因组中鉴定出FN3K同源物。在这些基因组中的11个基因组中,从系统发育远缘的细菌开始,FN3K同源物紧随其后是低分子量蛋白酪氨酸磷酸酶(LMW-PTP)同源物,因此,其功能可能与FN3K同源。在大肠杆菌中过表达了五种细菌FN3K同源物(来自大肠杆菌,粪肠球菌,植物乳杆菌,金黄色葡萄球菌和嗜热栖热菌),并对其纯化和动力学性质进行了研究。四种是核糖胺/赤藓糖胺3-激酶,对游离赖氨酸和尸胺衍生物作用最好,但对结合于氨基酸α-氨基的核糖胺作用最弱。它们还使结合蛋白的核糖胺或赤藓糖胺磷酸化,但不结合蛋白的果糖胺磷酸化,因此具有与哺乳动物FN3K相关蛋白相似的特性。大肠杆菌FN3K同源物(YniA)在所有受测底物上均无活性。嗜热链球菌的LMW-PTP与FN3K同源物形成操纵子,金黄色葡萄球菌的LMW-PTP(PtpA)在大肠杆菌中过表达,纯化并显示不仅使蛋白质酪氨酸磷酸酯而且使蛋白质脱磷酸化核糖胺5-磷酸酯以及游离的核糖赖氨酸5-磷酸酯和赤藓糖赖氨酸4-磷酸酯。这些LMW-PTP没有核糖胺3-磷酸酶活性。结论是大多数细菌FN3K同源物是核糖胺/赤藓糖胺3激酶。它们可以与磷酸酶一起用于使由5-磷酸核糖或4-磷酸赤糖形成的糖基化产物脱糖。

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