Misfolded forms of glyceraldehyde-3-phosphate dehydrogenase interact with GroEL and inhibit chaperonin-assisted folding of the wild-type enzyme.
We studied the interaction of chaperonin GroEL with different misfolded forms of tetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH): (1) GAPDH from rabbit muscles with all SH-groups modified by 5,5'-dithiobis(2-nitrobenzoate); (2) O-R-type dimers of mutant GAPDH from Bacillus stearothermophilus with amino acid substitutions Y283V, D282G, and Y283V/W84F, and (3) O-P-type dimers of mutant GAPDH from B. stearothermophilus with amino acid substitutions Y46G/S48G and Y46G/R52G. It was shown that chemically modified GAPDH and the O-R-type mutant dimers bound to GroEL with 1:1 stoichiometry and dissociation constants K(d) of 0.4 and 0.9 muM, respectively. A striking feature of the resulting complexes with GroEL was their stability in the presence of Mg-ATP. Chemically modified GAPDH and the O-R-type mutant dimers inhibited GroEL-assisted refolding of urea-denatured wild-type GAPDH from B. stearothermophilus but did not affect its spontaneous reactivation. In contrast to the O-R-dimers, the O-P-type mutant dimers neither bound nor affected GroEL-assisted refolding of the wild-type GAPDH. Thus, we suggest that interaction of GroEL with certain types of misfolded proteins can result in the formation of stable complexes and the impairment of chaperonin activity.
Polyakova
Oxana V
OV
Department of Biochemistry of Animal Cell, Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119992, Russian Federation.
Roitel
Olivier
O
Asryants
Regina A
RA
Poliakov
Alexei A
AA
Branlant
Guy
G
Muronetz
Vladimir I
VI
D000818:Animals; D018834:Chaperonin 60 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019281:Dimerization; D004228:Dithionitrobenzoic Acid / Q000737:chemistry; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D017510:Protein Folding; D011817:Rabbits; D013439:Sulfhydryl Reagents
D018834:Chaperonin 60; D013439:Sulfhydryl Reagents; D004228:Dithionitrobenzoic Acid; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Cell. 1999 Apr 30;97(3):325-38
10319813
Science. 1999 Apr 30;284(5415):822-5
10221918
Methods Enzymol. 1972;25:457-64
23014426
Science. 1999 Dec 3;286(5446):1888-93
10583944
Biochemistry. 1999 Dec 7;38(49):16084-91
10587431
Proc Natl Acad Sci U S A. 2000 May 23;97(11):5746-9
10811907
Biochemistry. 2002 Jun 18;41(24):7556-64
12056886
J Mol Cell Cardiol. 2002 Nov;34(11):1549-60
12431453
J Mol Biol. 2003 Mar 7;326(5):1513-22
12595262
Semin Cell Dev Biol. 2004 Feb;15(1):3-16
15036202
Nature. 1970 Aug 15;227(5259):680-5
5432063
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
388439
Biochim Biophys Acta. 1980 Jun 13;613(2):292-308
7004490
Methods Enzymol. 1982;90 Pt E:479-90
7154963
Anal Biochem. 1985 Dec;151(2):571-4
4096388
Biochemistry. 1991 Sep 24;30(38):9195-200
1680001
Biochemistry. 1991 Oct 8;30(40):9716-23
1680394
J Biol Chem. 1992 Sep 5;267(25):17773-9
1355477
Protein Sci. 1992 Mar;1(3):363-9
1363913
J Biol Chem. 1993 Sep 15;268(26):19346-51
8103517
J Biol Chem. 1994 May 6;269(18):13629-36
7909810
Science. 1994 Jul 29;265(5172):659-66
7913555
Nature. 1994 Oct 13;371(6498):578-86
7935790
J Biol Chem. 1995 Jan 20;270(3):1011-4
7836352
Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4509-12
8633099
Biochem J. 1996 May 1;315 ( Pt 3):1021-6
8645139
Protein Sci. 1996 Mar;5(3):478-87
8868484
Free Radic Biol Med. 1997;22(7):1277-82
9098102
Protein Sci. 1997 Apr;6(4):743-60
9098884
J Biol Chem. 1998 Jan 30;273(5):3027-32
9446617
Biochem J. 1998 Apr 15;331 ( Pt 2):505-11
9531491
Annu Rev Biochem. 1998;67:581-608
9759498
J Protein Chem. 1999 Jan;18(1):79-87
10071932
J Biol Chem. 1999 Apr 16;274(16):10790-4
10196153
Mech Ageing Dev. 1999 Mar 15;107(3):323-32
10360685