کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5913680 | 1162697 | 2015 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Structural insights on mouse l-threonine dehydrogenase: A regulatory role of Arg180 in catalysis
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
Nicotinamide adenine dinucleotide (NAD)medium-chain dehydrogenase/reductasel-threonine dehydrogenaseUDP-galactose 4-epimerasePRMT5mESCMTDHMDRSDRASUl-threonine - L-ترئونینGalE - بادtdh - بلهShort-chain dehydrogenase/reductase - دلائل زنجیره ای دهیدروژناز / ردوکتازDehydrogenase - دهیدروژنازCrystal structure - ساختار کریستالیMouse embryonic stem cell - سلول بنیادی جنینی موشEnzyme kinetics - سینتیک آنزیمMouse - موشwild-type - نوع وحشیasymmetric unit - واحد نامتقارنProtein arginine methyltransferase 5 - پروتئین آرژینین متیل ترانسفراز 5
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Mouse l-threonine dehydrogenase (mTDH), which belongs to the short-chain dehydrogenase/reductase (SDR) superfamily and mediates threonine catabolism, plays pivotal roles in both powerful biosynthesis and signaling in mouse stem cells and has a regulatory residue Arg180. Here we determined three crystal structures of mTDH: wild-type (WT) in the apo form; in complex with NAD+ and a substrate analog, glycerol, or with only NAD+; as well as the R180K variant with NAD+. This is the first description of a structure for mammalian SDR-type TDH. Structural comparison revealed the structural basis for SDR-type TDH catalysis remains strictly conserved in bacteria and mammals. Kinetic enzyme assays, and isothermal titration calorimetry (ITC) measurements indicated the R180K mutation has little effect on NAD+ binding affinity, whereas affects the substrate's affinity for the enzyme. The crystal structure of R180K with NAD+, biochemical and spectroscopic studies suggested that the R180K mutant should bind NAD+ in a similar way and have a similar folding to the WT. However, the R180K variant may have difficulty adopting the closed form due to reduced interaction of residue 180 with a loop which connects a key position for mTDH switching between the closed and open forms in mTDH catalysis, and thereby exhibited a significantly decreased kcat/Km value toward the substrate, l-Thr. In sum, our results suggest that activity of GalE-like TDH can be regulated by remote interaction, such as hydrogen bonding and hydrophobic interaction around the Arg180 of mTDH.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Structural Biology - Volume 192, Issue 3, December 2015, Pages 510-518
Journal: Journal of Structural Biology - Volume 192, Issue 3, December 2015, Pages 510-518
نویسندگان
Chao He, Xianyu Huang, Yanhong Liu, Fudong Li, Yang Yang, Hongru Tao, Chuanchun Han, Chen Zhao, Yazhong Xiao, Yunyu Shi,