Interactions among stalk modules of thrombospondin-1.
Thrombospondin-1 is a trimeric, modular calcium-binding glycoprotein. The subunit is composed of an N-terminal module; oligomerization domain; stalk modules including a von Willebrand factor type C module, three properdin or thrombospondin type 1 repeat (TSR) modules, and two thrombospondin-type EGF-like modules; and a C-terminal signature domain comprising single copies of the epidermal growth factor (EGF)-like, wire, and lectin-like modules. Conformational changes in the signature domain influence ligand binding to the N-terminal modules. Interactions have been demonstrated among the modules of the signature domain and the thrombospondin-type EGF-like modules. We have extended this analysis to the rest of the stalk modules. Differential scanning calorimetry revealed interactions between the most C-terminal TSR module and the EGF-like modules. Calorimetry and differences in expression levels of single versus tandem modules indicated that the three TSRs interact with each other as well. No evidence of interactions between the von Willebrand factor type C and TSR modules were detected by differential scanning calorimetry, circular dichroism, or intrinsic fluorescence. These results indicate that the TSR and thrombospondin-type EGF-like stalk modules act as a unit that may relay conformational information between the N-terminal and C-terminal parts of the protein.
Liu
Yuanyuan
Y
Department of Biomolecular Chemistry and Medicine, University of Wisconsin, Madison, Wisconsin 53706, USA.
Mosher
Deane F
DF
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000737:chemistry; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002942:Circular Dichroism / Q000379:methods; D003001:Cloning, Molecular; D004815:Epidermal Growth Factor / Q000737:chemistry; D006801:Humans; D007313:Insecta; D037102:Lectins / Q000737:chemistry; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods; D019700:Thrombospondin 1 / Q000737:chemistry / Q000378:metabolism*
D037102:Lectins; D011994:Recombinant Proteins; D019700:Thrombospondin 1; D004815:Epidermal Growth Factor; D002118:Calcium
J Biol Chem. 2001 Mar 2;276(9):6485-98
11067851
Methods Cell Biol. 2002;69:69-81
12071009
Structure. 2006 Jan;14(1):33-42
16407063
Matrix Biol. 2008 May;27(4):339-51
18226512
Science. 1966 Dec 9;154(3754):1288-99
5332570
J Biol Chem. 2008 Jul 18;283(29):20069-76
18499674
Curr Opin Cell Biol. 2000 Oct;12(5):634-40
10978901
J Thromb Haemost. 2006 Feb;4(2):459-68
16420580
J Biol Chem. 2004 Oct 1;279(40):41734-43
15292271
J Biol Chem. 2005 Dec 16;280(50):41229-35
16246837
J Biol Chem. 1984 Apr 10;259(7):4582-8
6707020
Structure. 2004 Dec;12(12):2173-83
15576031
J Cell Biol. 1985 Oct;101(4):1434-41
2413043
Methods Enzymol. 1990;182:50-68
2314256
Biochemistry. 1992 Feb 4;31(4):1173-80
1734965
J Biol Chem. 2004 May 14;279(20):21651-7
15007078
J Biol Chem. 2009 Aug 14;284(33):22206-22212
19531495
Anal Biochem. 1992 Jan;200(1):74-80
1595904
J Biochem. 1992 Apr;111(4):460-4
1618736
FASEB J. 2009 Aug;23(8):2490-501
19276170
Cell Mol Life Sci. 2008 Mar;65(5):728-42
18193160
Cell. 2008 Jan 25;132(2):247-58
18243100
J Cell Biol. 2002 Oct 28;159(2):373-82
12391027
Cell Mol Life Sci. 2008 Mar;65(5):672-86
18193164
Biochem Biophys Res Commun. 2002 Aug 9;296(1):156-60
12147243
Cell Mol Life Sci. 2008 Mar;65(5):713-27
18193161
J Mol Biol. 2004 Nov 5;343(5):1327-43
15491616
Biochemistry. 2002 Dec 3;41(48):14329-39
12450399
J Biol Chem. 2004 Dec 17;279(51):53857-66
15466413
J Biol Chem. 2000 Dec 29;275(52):40938-45
11016937
EMBO J. 2006 Nov 15;25(22):5270-83
17082774
Biochemistry. 1988 Aug 23;27(17):6214-20
3219333
Nat Struct Mol Biol. 2005 Oct;12(10):910-4
16186819
Nat Med. 2000 Jan;6(1):41-8
10613822
Annu Rev Biochem. 2004;73:1019-49
15189166
J Biol Chem. 2007 Jun 8;282(23):17024-31
17395588
J Biol Chem. 2004 Nov 12;279(46):47633-42
15342643
J Biol Chem. 1985 Mar 25;260(6):3762-72
2579080
J Biol Chem. 2007 Jun 8;282(23):17014-23
17395589
J Biol Chem. 2008 Feb 15;283(7):3932-41
18065761
J Biol Chem. 1984 Mar 25;259(6):3944-8
6231289
Dev Cell. 2008 May;14(5):739-50
18477456
J Biol Chem. 2007 Sep 14;282(37):27067-27075
17620335