Reducing the effects of intracellular accumulation of thermolabile collagen II mutants by increasing their thermostability in cell culture conditions.
Mutations in collagen II are associated with spondyloepiphyseal dysplasia, a group of heritable diseases whose common features include aberrations of skeletal growth. The mechanisms through which mutations in collagen II affect the cartilaginous tissues are complex and include both intracellular and extracellular processes. One of those mechanisms involves cellular stress caused by excessive accumulation of misfolded collagen II mutants. We investigated whether stabilizing the structure of thermolabile R789C and R992C collagen II mutants would improve their secretion from cells, thereby reducing cellular stress and apoptosis. Employing glycerol and trimethylamine N-oxide (TMAO), chemicals that increase the thermostability of collagen triple helices, we demonstrated that those compounds function as chaperones and stabilize the R789C and R992C mutants, accelerate their secretion, and improve cell survival. Our study provides a scientific basis for considering misfolded triple helices of collagen mutants a target for reducing the deleterious effects caused by their excessive intracellular accumulation.
Gawron
Katarzyna
K
Department of Dermatology and Cutaneous Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Jensen
Deborah A
DA
Steplewski
Andrzej
A
Fertala
Andrzej
A
D017209:Apoptosis; D018929:Cell Culture Techniques; D024043:Collagen Type II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005990:Glycerol / Q000737:chemistry / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D008744:Methylamines / Q000737:chemistry / Q000494:pharmacology; D009154:Mutation; D010009:Osteochondrodysplasias / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D055550:Protein Stability
D024043:Collagen Type II; D008744:Methylamines; C005855:trimethyloxamine; D005990:Glycerol
Proc Natl Acad Sci U S A. 1981 Aug;78(8):5142-6
6946461
Br J Dermatol. 2008 Jul;159(1):252-5
18489603
Bone. 1995 Aug;17(2 Suppl):45S-49S
8579897
Pediatr Res. 2002 Dec;52(6):832-6
12438657
Biochim Biophys Acta. 2006 Nov;1764(11):1677-87
17046342
J Protein Chem. 1999 May;18(4):397-401
10449037
Biochem J. 1973 Feb;131(2):335-42
4737319
Annu Rev Biochem. 1995;64:403-34
7574488
Nat Rev Drug Discov. 2008 Dec;7(12):1013-30
19043451
Swiss Med Wkly. 2002 May 4;132(17-18):211-22
12087487
J Bioenerg Biomembr. 1997 Oct;29(5):491-502
9511934
Nature. 2003 Dec 18;426(6968):905-9
14685252
J Mol Biol. 2005 Sep 16;352(2):382-95
16083907
J Biol Chem. 2001 Oct 26;276(43):40241-6
11507102
J Biol Chem. 1993 Oct 5;268(28):21232-7
8407960
Exp Cell Res. 2008 Mar 10;314(5):950-60
18255062
J Biol Chem. 1979 Apr 10;254(7):2234-43
570970
J Mol Biol. 2004 May 14;338(5):989-98
15111062
Ann N Y Acad Sci. 2003 May;995:140-50
12814946
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4273-8
10725403
Science. 2006 Aug 25;313(5790):1137-40
16931765
J Mol Biol. 2009 Jul 10;390(2):306-18
19433093
J Biol Chem. 1998 Feb 27;273(9):4831-4
9478922
Hum Mutat. 2008 Jun;29(6):841-51
18383211
J Biol Chem. 1993 Jul 15;268(20):15238-45
8325895
Annu Rev Biochem. 1974;43(0):567-603
4605221
J Biol Chem. 1983 May 10;258(9):5915-21
6304100
Biochem J. 1994 Feb 15;298 ( Pt 1):31-7
8129728
J Bone Miner Res. 2003 Sep;18(9):1612-21
12968670
Arch Biochem Biophys. 2005 Apr 1;436(1):78-82
15752711
Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1796-801
10677536