Order within disorder: aggrecan chondroitin sulphate-attachment region provides new structural insights into protein sequences classified as disordered.
Structural investigation of proteins containing large stretches of sequences without predicted secondary structure is the focus of much increased attention. Here, we have produced an unglycosylated 30 kDa peptide from the chondroitin sulphate (CS)-attachment region of human aggrecan (CS-peptide), which was predicted to be intrinsically disordered and compared its structure with the adjacent aggrecan G3 domain. Biophysical analyses, including analytical ultracentrifugation, light scattering, and circular dichroism showed that the CS-peptide had an elongated and stiffened conformation in contrast to the globular G3 domain. The results suggested that it contained significant secondary structure, which was sensitive to urea, and we propose that the CS-peptide forms an elongated wormlike molecule based on a dynamic range of energetically equivalent secondary structures stabilized by hydrogen bonds. The dimensions of the structure predicted from small-angle X-ray scattering analysis were compatible with EM images of fully glycosylated aggrecan and a partly glycosylated aggrecan CS2-G3 construct. The semiordered structure identified in CS-peptide was not predicted by common structural algorithms and identified a potentially distinct class of semiordered structure within sequences currently identified as disordered. Sequence comparisons suggested some evidence for comparable structures in proteins encoded by other genes (PRG4, MUC5B, and CBP). The function of these semiordered sequences may serve to spatially position attached folded modules and/or to present polypeptides for modification, such as glycosylation, and to provide templates for the multiple pleiotropic interactions proposed for disordered proteins.
Jowitt
Thomas A
TA
Wellcome Trust Centre for Cell Matrix Research, University of Manchester, Manchester, M13 9PT, United Kingdom. t.jowitt@manchester.ac.uk
Murdoch
Alan D
AD
Baldock
Clair
C
Berry
Richard
R
Day
Joanna M
JM
Hardingham
Timothy E
TE
D053674:Aggrecans / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001665:Binding Sites; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000378:metabolism*; D019295:Computational Biology; D006801:Humans; D057446:Hydrodynamics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011489:Protein Denaturation / Q000187:drug effects; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D053838:Scattering, Small Angle; D040681:Structural Homology, Protein; D014508:Urea / Q000494:pharmacology; D014961:X-Ray Diffraction
D053674:Aggrecans; D010455:Peptides; D014508:Urea; D002809:Chondroitin Sulfates
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