title: Biophysical and structural characterization of a folded core domain within the proregion of growth and differentiation factor-5.
abstract: The structure and function(s) of the very large proregions of the transforming growth factor-β structure family are known in only a few cases. The proregion of growth and differentiation factor (GDF)5 comprises 354 residues. GDF5 therefore belongs to the group of those growth factors with the largest proregions. Here, we report a biophysical analysis of the proform (proGDF5) and the separate proregion. In the absence of the mature part, the proregion folds reversibly to form a monomeric polypeptide that is stabilized by an intramolecular disulfide bond. In the context of the mature part, i.e. in proGDF5, the proregion shows increased thermodynamic stability and contains a higher proportion of secondary structural elements than in its isolated form. A subdomain within the proregion represents a well-folded structure as monitored via biophysical analysis and NMR spectroscopy. Furthermore, two point mutations that are associated with skeletal malformations lead to reduced thermodynamic stability, which is interpreted on the basis of a homology model with the structure of the related latency-associated peptide, representing the proregion of transforming growth factor-β1.
authors.affiliation: Technical Biochemistry, Martin-Luther-University Halle-Wittenberg, Germany.
authors.forename: Tino
authors.initials: T
authors.lastname: Thieme
authors.forename: Rica
authors.initials: R
authors.lastname: Patzschke
authors.forename: Florian
authors.initials: F
authors.lastname: Job
authors.forename: Jens
authors.initials: J
authors.lastname: Liebold
authors.forename: Petra
authors.initials: P
authors.lastname: Seemann
authors.forename: Hauke
authors.initials: H
authors.lastname: Lilie
authors.forename: Jochen
authors.initials: J
authors.lastname: Balbach
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Schwarz
mesh_terms: D019943:Amino Acid Substitution; D059327:Brachydactyly / Q000235:genetics; D002942:Circular Dichroism; D003553:Cystine / Q000032:analysis; D055428:Growth Differentiation Factor 5 / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology; D006358:Hot Temperature; D006801:Humans; D008958:Models, Molecular; D020125:Mutation, Missense; D019906:Nuclear Magnetic Resonance, Biomolecular; D017354:Point Mutation; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
chemical_list: C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D003553:Cystine
keywords: bone morphogenetic protein; domain coupling; proregion; subdomain; thermodynamics