Effect of Detergents on Galactoside Binding by Melibiose Permeases

Biochemistry
Anowarul AminLan Guan

Abstract

The effect of various detergents on the stability and function of the melibiose permeases of Escherichia coli (MelBEc) and Salmonella typhimurium (MelBSt) was studied. In n-dodecyl-β-d-maltoside (DDM) or n-undecyl-β-d-maltoside (UDM), WT MelBSt binds melibiose with an affinity similar to that in the membrane. However, with WT MelBEc or MelBSt mutants (Arg141 → Cys, Arg295 → Cys, or Arg363 → Cys), galactoside binding is not detected in these detergents, but binding to the phosphotransferase protein IIA(Glc) is maintained. In the amphiphiles lauryl maltose neopentyl glycol (MNG-3) or glyco-diosgenin (GDN), galactoside binding with all of the MelB proteins is observed, with slightly reduced affinities. MelBSt is more thermostable than MelBEc, and the thermostability of either MelB is largely increased in MNG-3 or GDN. Therefore, the functional defect with DDM or UDM likely results from the relative instability of the sensitive MelB proteins, and stability, as well as galactoside binding, is retained in MNG-3 or GDN. Furthermore, isothermal titration calorimetry of melibiose binding with MelBSt shows that the favorable entropic contribution to the binding free energy is decreased in MNG-3, indicating that the conformational dynamic...Continue Reading

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Citations

Apr 26, 2016·Chemical Science·Aiman SadafPil Seok Chae
Mar 12, 2016·Journal of the American Chemical Society·Muhammad EhsanPil Seok Chae
Oct 22, 2017·The Journal of General Physiology·Parameswaran Hariharan, Lan Guan
Apr 6, 2018·Chemical Science·Aiman SadafPil Seok Chae
Apr 17, 2018·Chembiochem : a European Journal of Chemical Biology·Muhammad EhsanPil Seok Chae
Jun 24, 2017·Scientific Reports·Hazrat HussainPil Seok Chae
Jun 11, 2021·The Journal of General Physiology·Parameswaran Hariharan, Lan Guan
Dec 15, 2020·Journal of the American Chemical Society·Manabendra DasPil Seok Chae
Jun 6, 2020·ACS Chemical Biology·Muhammad EhsanPil Seok Chae
Dec 7, 2019·Journal of the American Chemical Society·Lubna GhaniPil Seok Chae

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