首页> 外文OA文献 >The skin of atopic dermatitis patients contains a novel enzyme, glucosylceramide sphingomyelin deacylase, which cleaves the N-acyl linkage of sphingomyelin and glucosylceramide.
【2h】

The skin of atopic dermatitis patients contains a novel enzyme, glucosylceramide sphingomyelin deacylase, which cleaves the N-acyl linkage of sphingomyelin and glucosylceramide.

机译:特应性皮炎患者的皮肤含有一种新型酶,葡糖神经酰胺鞘磷脂脱酰基酶,该酶可裂解鞘磷脂和葡糖神经酰胺的N-酰基键。

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务,本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文,但由于OA文献来源多样且变更频繁,仍可能出现获取不到、文献不完整或与标题不符等情况,如果获取不到我们将提供退款服务。请知悉。

摘要

We have demonstrated previously that there is an abnormal expression of sphingomyelin (SM) deacylase in the epidermis of patients with atopic dermatitis (ADe). In the present study, we have prepared N-[palmitic acid-1-(14)C]SM and N-[palmitic acid-1-(14)C]glucosylceramide (GCer) to use as substrates and have quantified SM deacylase activity by detecting the release of [(14)C]palmitic acid in extracts of the stratum corneum or the epidermis of ADe patients. In studies using [palmitic acid-1-(14)C]SM as a substrate, a pH dependency of catalytic activity with a peak at pH 5.0 was found. Preparative SDS/PAGE using an extract of ADe epidermis revealed that the molecular mass of SM deacylase is 40000 Da, which is consistent with its apparent molecular mass of 42000 Da estimated by gel-filtration analysis of stratum corneum extracts. Analytical isoelectric focusing (IEF) chromatography demonstrated that the pI values of SM deacylase, beta-glucocerebrosidase (GlcCDase), sphingomyelinase (SMase) and acid ceramidase were 4.2, 7.4, 7.0 and 5.7, respectively. In enzymic analysis using pI-4.2 SM deacylase partially purified by IEF, which had no detectable contamination with acid ceramidase, GlcCDase or SMase, radio-TLC analysis revealed that radiolabelled sphingosylphosphocholine or [1-(14)C]palmitic acid was enzymically liberated from [choline-methyl-(14)C]SM or N-[palmitoyl-1-(14)C]GCer, respectively, used as substrates. Further the pI-4.2 protein purified from extracts of the stratum corneum of ADe patients was able to hydrolyse N-[palmitoyl-1-(14)C]SM and GCer, but not N-[palmitoyl-1-(14)C]ceramide. These results indicate that a hitherto undiscovered epidermal enzyme, termed here glucosylceramide sphingomyelin deacylase, is expressed in the skin of ADe patients, which plays an important role in ceramide deficiency (including acylceramides) in the stratum corneum.
机译:以前我们已经证明特应性皮炎(ADe)患者的表皮中鞘磷脂(SM)脱酰基酶表达异常。在本研究中,我们制备了N- [棕榈酸-1-(14)C] SM和N- [棕榈酸-1-(14)C]葡萄糖基神经酰胺(GCer)作为底物并定量了SM脱酰基酶活性通过检测ADe患者的角质层或表皮提取物中[(14)C]棕榈酸的释放。在使用[棕榈酸-1-(14)C] SM作为底物的研究中,发现了催化活性的pH依赖性,其峰值在pH 5.0。使用ADe表皮提取物进行的制备SDS / PAGE显示,SM脱酰基酶的分子量为40000 Da,这与通过角质层提取物的凝胶过滤分析估计的42000 Da的表观分子量相符。等电聚焦分析(IEF)色谱表明,SM脱酰基酶,β-葡萄糖脑苷脂酶(GlcCDase),鞘磷脂酶(SMase)和酸性神经酰胺酶的pI值分别为4.2、7.4、7.0和5.7。在使用经IEF部分纯化的pI-4.2 SM脱酰基酶的酶分析中,酸的神经酰胺酶,GlcCDase或SMase均未检测到污染,放射性TLC分析表明放射性标记的鞘氨醇磷酸胆碱或[1-(14)C]棕榈酸是从酶中释放的分别使用[胆碱-甲基-(14)C] SM或N- [棕榈酰基-1-(14)C] GCer作为底物。此外,从ADe患者角质层提取物中纯化的pI-4.2蛋白能够水解N- [palmitoyl-1-(14)C] SM和GCer,但不能水解N- [palmitoyl-1-(14)C]神经酰胺。这些结果表明,迄今尚未发现的表皮酶,在这里被称为葡萄糖基神经酰胺鞘磷脂脱酰基酶,在ADe患者的皮肤中表达,其在角质层中的神经酰胺缺乏(包括酰基神经酰胺)中起重要作用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号