In Porphyromonas gingivalis VimF is involved in gingipain maturation through the transfer of galactose.
In Porphyromonas gingivalis VimF is involved in gingipain maturation through the transfer of galactose.
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DOI：
10.1371/journal.pone.0063367
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Fletcher HM
中科院分区：
文献类型：
--
作者：
Muthiah AS;Aruni W;Robles AG;Dou Y;Roy F;Fletcher HM
在此之前，我们已经报道了牙龈卟啉单胞菌中的牙龈痛活性是由包括VimF在内的独特的Vim蛋白翻译后调节的，VimF是一种可能的糖基转移酶。为了进一步确定VimF的特征，我们评估了一个在不同牙龈假单胞菌遗传背景中该基因缺陷的等位基因突变体。此外，用重组VimF蛋白进一步证实了其糖基转移酶功能。33277菌株遗传背景中的突变株(FLL476)的表型与W83菌株中的突变株(FLL95)相似。在未检测到血凝性和自聚集性降低的情况下，FLL476生物膜的形成增加。用牙龈假单胞菌FLL95和FLL476孵育的HeLa细胞侵袭能力下降了45%。在大肠杆菌中产生的针对重组VimF蛋白的抗体只与牙龈假单胞菌脱糖的天然VimF蛋白发生免疫反应。分别以UDP-半乳糖和N-乙酰氨基葡萄糖为供体和受体底物，观察rVimF的体外糖基转移酶活性。在rVimF和UDP-半乳糖存在下，经质谱仪鉴定为RGP牙龈痛的FLL95胞外部分的60 kDa蛋白与糖链特异性mAb1B5抗体发生免疫反应。综上所述，这些结果表明VimF糖蛋白是一种半乳糖基转移酶，可能是牙龈痛糖基化所特有的。此外，半乳糖对不断增长的糖链至关重要。
Previously, we have reported that gingipain activity in Porphyromonas gingivalis, the major causative agent in adult periodontitis, is post-translationally regulated by the unique Vim proteins including VimF, a putative glycosyltransferase. To further characterize VimF, an isogenic mutant defective in this gene in a different P. gingivalis genetic background was evaluated. In addition, the recombinant VimF protein was used to further confirm its glycosyltransferase function. The vimF-defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF-defective mutant (FLL95) in the P. gingivalis W83 genetic background. While hemagglutination was not detected and autoaggregation was reduced, biofilm formation was increased in FLL476. HeLa cells incubated with P. gingivalis FLL95 and FLL476 showed a 45% decrease in their invasive capacity. Antibodies raised against the recombinant VimF protein in E. coli immunoreacted only with the deglycosylated native VimF protein from P. gingivalis. In vitro glycosyltransferase activity for rVimF was observed using UDP-galactose and N-acetylglucosamine as donor and acceptor substrates, respectively. In the presence of rVimF and UDP-galactose, a 60 kDa protein from the extracellular fraction of FLL95 which was identified by mass spectrometry as Rgp gingipain, immunoreacted with the glycan specific mAb 1B5 antibody. Taken together, these results suggest the VimF glycoprotein is a galactosyltransferase that may be specific for gingipain glycosylation. Moreover, galatose is vital for the growing glycan chain.
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影响因子：
2.9
作者：
Larkin A;Imperiali B
通讯作者：
Imperiali B
影响因子：
3.1
作者：
Abaibou, H;Chen, Z;Fletcher, HM
通讯作者：
Fletcher, HM
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作者：
Aruni, A. Wilson;Lee, J.;Fletcher, H. M.
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Fletcher, H. M.
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Davies, GJ;Gloster, TM;Henrissat, B
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Henrissat, B
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作者：
Gross, Julia;Grass, Susan;Geme, Joseph W. St., III
通讯作者：
Geme, Joseph W. St., III