Location of 3-Hydroxyproline Residues in Collagen Types I, II, III, and V/XI Implies a Role in Fibril Supramolecular Assembly

• Published in: The Journal of biological chemistry 2010-01, Vol.285 (4), p.2580-2590
• Main Authors: Weis, Mary Ann, Hudson, David M., Kim, Lammy, Scott, Melissa, Wu, Jiann-Jiu, Eyre, David R.
• Format: Article
• Language: English
• Subjects: 3-hydroxyproline; Adult; Amino Acid Sequence; Animals; Bone; Bone and Bones - chemistry; Bone and Bones - metabolism; Cartilage; Cartilage - chemistry; Cartilage - metabolism; Cattle; Chickens; Collagen - chemistry; Collagen - genetics; Collagen - metabolism; Collagen Type I - chemistry; Collagen Type I - genetics; Collagen Type I - metabolism; Collagen Type II - chemistry; Collagen Type II - genetics; Collagen Type II - metabolism; Collagen Type III - chemistry; Collagen Type III - genetics; Collagen Type III - metabolism; Collagen Type V - chemistry; Collagen Type V - genetics; Collagen Type V - metabolism; Collagen Type XI - chemistry; Collagen Type XI - genetics; Collagen Type XI - metabolism; Extracellular Matrix Proteins - chemistry; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Extracellular Matrix/Collagen; Extracellular Matrix/Hydroxyproline; Humans; Hydroxyproline - chemistry; Hydroxyproline - genetics; Hydroxyproline - metabolism; Molecular Sequence Data; Organisms/Mammal; Protein Processing, Post-Translational; Protein Structure and Folding; Protein/Post-translational Modification; Tandem Mass Spectrometry; Young Adult
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M109.068726

Collagen triple helices are stabilized by 4-hydroxyproline residues. No function is known for the much less common 3-hydroxyproline (3Hyp), although genetic defects inhibiting its formation cause recessive osteogenesis imperfecta. To help understand the pathogenesis, we used mass spectrometry to identify the sites and local sequence motifs of 3Hyp residues in fibril-forming collagens from normal human and bovine tissues. The results confirm a single, essentially fully occupied 3Hyp site (A1) at Pro986 in A-clade chains α1(I), α1(II), and α2(V). Two partially modified sites (A2 and A3) were found at Pro944 in α1(II) and α2(V) and Pro707 in α2(I) and α2(V), which differed from A1 in sequence motif. Significantly, the distance between sites 2 and 3, 237 residues, is close to the collagen D-period (234 residues). A search for additional D-periodic 3Hyp sites revealed a fourth site (A4) at Pro470 in α2(V), 237 residues N-terminal to site 3. In contrast, human and bovine type III collagen contained no 3Hyp at any site, despite a candidate proline residue and recognizable A1 sequence motif. A conserved histidine in mammalian α1(III) at A1 may have prevented 3-hydroxylation because this site in chicken type III was fully hydroxylated, and tyrosine replaced histidine. All three B-clade type V/XI collagen chains revealed the same three sites of 3Hyp but at different loci and sequence contexts from those in A-clade collagen chains. Two of these B-clade sites were spaced apart by 231 residues. From these and other observations we propose a fundamental role for 3Hyp residues in the ordered self-assembly of collagen supramolecular structures.