کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5584288 | 1567894 | 2017 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Multiple exo-glycosidases in human serum as detected with the substrate DNP-α-GalNAc. I. A new assay for lysosomal α-N-acetylgalactosaminidase
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کلمات کلیدی
DMFDNPHA380GLA4-methylumbelliferoneS.A.2,4-Dinitrophenol - 2،4-دینیتروفنلDMSO - DMSOα-N-acetylgalactosaminidase - α-N-استیل گلاکتوزامینیدازα-Galactosidase A - α-گالاکتوزیداز ANaga - اژدهاRoom temperature - دمای اتاقdimethylsulphoxide - دی متیل سولفوکسیدdimethylformamide - دی متیل فرمالیدLysosomes - لیزوزوم ها
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوشیمی بالینی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
This paper presents a new assay to determine the activity of the lysosomal enzyme α-N-acetylgalactosaminidase (Naga, EC 3.2.1.49) in human serum. It is based on the use of a new chromogenic substrate, DNP-α-GalNAc (2,4-dinitrophenyl-N-acetyl-α-D-galactosaminide) and is performed at pH 4.3 and 37 °C. This allows continuous monitoring of the absorbance of the released DNP. The assay can be performed with a standard spectrophotometer. Compared to established methods using an endpoint assay with MU-α-GalNAc (4-methylumbelliferyl-GalNAc), the present method gives a ca. 3-fold higher specific activity, while only one tenth of the serum concentration in the assay is required. Hence, the assay is at least 30-fold more sensitive than that with MU-α-GalNAc. The pH dependence of the reaction with DNP-α-GalNAc in the pH 3.5 to 6.5 region, while using 4% serum in the assay, shows only one peak around pH 4. This pH optimum is similar to that reported with MU-α-GalNAc. In the accompanying paper (S.P.J Albracht and J. Van Pelt (2017) Multiple exo-glycosidases in human serum as detected with the substrate DNP-α-GalNAc. II. Three α-N-acetylgalactosaminidase-like activities in the pH 5 to 8 region. BBA Clin. 8 (2017) 90-96), the method is used to show that, under special assay conditions, three more Naga-like activities can be uncovered in human serum.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: BBA Clinical - Volume 8, December 2017, Pages 84-89
Journal: BBA Clinical - Volume 8, December 2017, Pages 84-89
نویسندگان
Simon P.J. Albracht, Erik Allon, Johannes van Pelt,