title: Vascular Ehlers-Danlos syndrome mutations in type III collagen differently stall the triple helical folding.
abstract: Vascular Ehlers-Danlos syndrome (EDS) type IV is the most severe form of EDS. In many cases the disease is caused by a point mutation of Gly in type III collagen. A slower folding of the collagen helix is a potential cause for over-modifications. However, little is known about the rate of folding of type III collagen in patients with EDS. To understand the molecular mechanism of the effect of mutations, a system was developed for bacterial production of homotrimeric model polypeptides. The C-terminal quarter, 252 residues, of the natural human type III collagen was attached to (GPP)7 with the type XIX collagen trimerization domain (NC2). The natural collagen domain forms a triple helical structure without 4-hydroxylation of proline at a low temperature. At 33 °C, the natural collagenous part is denatured, but the C-terminal (GPP)7-NC2 remains intact. Switching to a low temperature triggers the folding of the type III collagen domain in a zipper-like fashion that resembles the natural process. We used this system for the two known EDS mutations (Gly-to-Val) in the middle at Gly-910 and at the C terminus at Gly-1018. In addition, wild-type and Gly-to-Ala mutants were made. The mutations significantly slow down the overall rate of triple helix formation. The effect of the Gly-to-Val mutation is much more severe compared with Gly-to-Ala. This is the first report on the folding of collagen with EDS mutations, which demonstrates local delays in the triple helix propagation around the mutated residue.
authors: Shriners Hospitals for Children Portland Research Center, Portland, Oregon 97239, USA.
authors: Kazunori
authors: K
authors: Mizuno
authors: Sergei
authors: S
authors: Boudko
authors: Jürgen
authors: J
authors: Engel
authors: Hans Peter
authors: HP
authors: Bächinger
mesh_terms: D002942:Circular Dichroism; D003094:Collagen / Q000737:chemistry; D024061:Collagen Type III / Q000737:chemistry / Q000235:genetics*; D004535:Ehlers-Danlos Syndrome / Q000235:genetics*; D006801:Humans; D009154:Mutation; D019696:Peptidylprolyl Isomerase / Q000737:chemistry; D017354:Point Mutation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D014357:Trypsin / Q000737:chemistry
chemical_list: D024061:Collagen Type III; D011994:Recombinant Proteins; D003094:Collagen; D014357:Trypsin; D019696:Peptidylprolyl Isomerase
keywords: Collagen; Connective Tissue; Ehlers-Danlos Syndrome; Glycine Mutations; Mutant; Prolyl Isomerase; Protein Folding; Type III Collagen
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