کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10796142 | 1052695 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Multiple proteins with single activities or a single protein with multiple activities: The conundrum of cell surface NADH oxidoreductases
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کلمات کلیدی
PBSCNOXtNOXMPMsWST-1pCMBSgp96BN-PAGEpreimmunePMSFHBSSNBTNADH oxidasePVDFFBSp-chloromercuriphenylsulfonic acidKCN - IP هاSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدSDS polyacrylamide gel electrophoresis - الکتروفورز ژل پلی آکریل آمید SDSblue native polyacrylamide gel electrophoresis - الکتروفورز ژل پلی آکریل آمید آبی بومیnitro blue tetrazolium - تترازولیم نیترو آبیRoom temperature - دمای اتاقpolyvinylidene difluoride - دی فلوئورید پلی وینیلیدینfoetal bovine serum - سرم جنین گاوPlasma membrane - غشای پلاسماPhosphate buffered saline - فسفات بافر شورphenylmethylsulfonyl fluoride - فنیل متیل سولفونیل فلورایدpotassium cyanide - پتاسیم سیانیدCapsaicin - کپسایسین یا کاپسیسین
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
دانش گیاه شناسی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Multiple proteins with single activities or a single protein with multiple activities: The conundrum of cell surface NADH oxidoreductases Multiple proteins with single activities or a single protein with multiple activities: The conundrum of cell surface NADH oxidoreductases](/preview/png/10796142.png)
چکیده انگلیسی
Reduction of the cell-impermeable tetrazolium salt WST-1 has been used to characterise two plasma membrane NADH oxidoreductase activities in human cells. The trans activity, measured with WST-1 and the intermediate electron acceptor mPMS, utilises reducing equivalents from intracellular sources, while the surface activity, measured with WST-1 and extracellular NADH, is independent of intracellular metabolism. Whether these two activities involve distinct proteins or are inherent to a single protein is unclear. In this work, we have attempted to address this question by examining the relationship between the trans and surface WST-1-reducing activities and a third well-characterised family of cell surface oxidases, the ECTO-NOX proteins. Using blue native-polyacrylamide gel electrophoresis, we have identified a complex in the plasma membranes of human 143B osteosarcoma cells responsible for the NADH-dependent reduction of WST-1. The dye-reducing activity of the 300 kDa complex was attributed to a 70 kDa NADH oxidoreductase activity that cross-reacted with antisera against the ECTO-NOX protein CNOX. Differences in enzyme activities and inhibitor profiles between the WST-1-reducing NADH oxidoreductase enzyme in the presence of NADH or mPMS and the ECTO-NOX family are reconciled in terms of the different purification methods and assay systems used to study these proteins.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Bioenergetics - Volume 1708, Issue 1, 1 June 2005, Pages 108-119
Journal: Biochimica et Biophysica Acta (BBA) - Bioenergetics - Volume 1708, Issue 1, 1 June 2005, Pages 108-119
نویسندگان
Debbie-Jane G. Scarlett, Patries M. Herst, Michael V. Berridge,