A Single Amino Acid Substitution Makes WNK4 Susceptible to SB 203580 and SB 202190.
Regulation of the SLC12 family of membrane transporters including NCCT involves a scaffold of interacting proteins including the STE 20 kinase SPAK and the WNK kinases, WNK 1 and WNK 4, which are mutated in the hypertensive syndrome of pseudohypoaldosteronism type 2 (PHAII). WNK4 regulates NCCT by affecting forward trafficking to the surface membrane. Studies in Xenopus using kinase dead WNK4 site mutants have produced inconsistent results with regard to the necessity of kinase function for NCCT regulation. Dynamic inhibition of WNK4 by small molecules may bring clarity to this issue however WNK4 is naturally resistant to commercial MAP kinase inhibitors owing to steric constraints prohibiting entry of small molecules to the active site. Using an approach similar to that used in p38 and ERK, we show that a single substitution in WNK4 (T261G) dramatically enhances its susceptibility to the inhibitors SB 202190 and SB 203580.
Clinical Pharmacology Unit, Department of Medicine, University of Cambridge, Cambridge, UK.
Mark
M
Glover
Connor
C
Sweeny
Bill
B
Davis
Kevin M
KM
O'Shaughnessy
J Am Soc Nephrol. 2003 Oct;14(10):2447-56
14514722
EMBO J. 1998 Aug 3;17(15):4426-41
9687510
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14109-14
14610273
Protein Sci. 1998 Nov;7(11):2249-55
9827991
Science. 2001 Aug 10;293(5532):1107-12
11498583
Proc Natl Acad Sci U S A. 2005 Jul 19;102(29):10315-20
16006511
Hypertension. 2005 Aug;46(2):295-300
15998707
Pharmacol Ther. 2005 May;106(2):221-31
15866321
Oncogene. 2008 Sep 18;27(42):5648-50
18504432
Nat Struct Biol. 1997 Apr;4(4):311-6
9095200
J Hypertens. 2001 Nov;19(11):1957-64
11677360
J Clin Invest. 2005 May;115(5):1379-87
15841204
Mod Rheumatol. 2008;18(4):366-78
18437285
Biol Pharm Bull. 2007 Sep;30(9):1610-6
17827708
Trends Cell Biol. 1997 Sep;7(9):353-61
17708980
J Am Soc Nephrol. 2006 Sep;17(9):2402-13
16899520
Chem Biol. 1998 Jun;5(6):321-8
9653550
Lancet. 2000 Jul 15;356(9225):194-202
10963197
J Biol Chem. 1994 Jul 1;269(26):17713-22
8021284
Am J Physiol Renal Physiol. 2006 Dec;291(6):F1369-76
16788137
N Engl J Med. 2003 Mar 13;348(11):994-1004
12637609
Methods Mol Biol. 2000;99:161-75
10909084
Yakugaku Zasshi. 2007 Mar;127(3):501-14
17329936
Nature. 1994 Dec 22-29;372(6508):739-46
7997261
EMBO J. 1998 Jun 15;17(12):3372-84
9628874
Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):680-4
12515852