title: Interactions among stalk modules of thrombospondin-1.
abstract: 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.
authors.affiliation: Department of Biomolecular Chemistry and Medicine, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Deane F
authors.initials: DF
authors.lastname: Mosher
mesh_terms: 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*
chemical_list: D037102:Lectins; D011994:Recombinant Proteins; D019700:Thrombospondin 1; D004815:Epidermal Growth Factor; D002118:Calcium
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