1. Type I and type V procollagen triple helix uses different subsets of the molecular ensemble for lysine posttranslational modifications in the rER
- Author
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Sara F. Tufa, Johanna Myllyharju, Douglas R. Keene, Kazunori Mizuno, Paul Holden, Hans Peter Bächinger, Douglas B. Gould, Olesya Semenova, Yuki Taga, Nobuyo Mizuno, Keith D. Zientek, Yoshihiro Ishikawa, and Antti M. Salo
- Subjects
Male ,collagen ,0301 basic medicine ,Protein Conformation ,Lysine ,CRTAP, cartilage-associated protein ,lysyl hydroxylase ,rER, rough endoplasmic reticulum ,GGHL, glucosylgalactosyl hydroxylysine ,Hydroxylysine ,Biochemistry ,Hydroxylation ,Mice ,chemistry.chemical_compound ,SFM, serum-free media ,Mice, Knockout ,AAA, amino acid analysis ,biology ,Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase ,molecular chaperone ,ECM, extracellular matrix ,endoplasmic reticulum ,prolyl hydroxylase ,posttranslational modifications ,Endoplasmic Reticulum, Rough ,Type I collagen ,Research Article ,Triple helix ,Lysyl hydroxylase ,Procollagen-Proline Dioxygenase ,Collagen Type I ,03 medical and health sciences ,CypB, cyclophilin B ,Animals ,EDS, Ehlers–Danlos syndrome ,Molecular Biology ,LH 1, lysyl hydroxylase 1 ,030102 biochemistry & molecular biology ,Endoplasmic reticulum ,Cell Biology ,Mice, Inbred C57BL ,Procollagen peptidase ,030104 developmental biology ,chemistry ,biology.protein ,P3H3, prolyl 3-hydroxylase 3 ,PTM, posttranslational modification ,Collagen Type V ,Protein Processing, Post-Translational - Abstract
Collagen is the most abundant protein in humans. It has a characteristic triple-helix structure and is heavily posttranslationally modified. The complex biosynthesis of collagen involves processing by many enzymes and chaperones in the rough endoplasmic reticulum. Lysyl hydroxylase 1 (LH1) is required to hydroxylate lysine for cross-linking and carbohydrate attachment within collagen triple helical sequences. Additionally, a recent study of prolyl 3-hydroxylase 3 (P3H3) demonstrated that this enzyme may be critical for LH1 activity; however, the details surrounding its involvement remain unclear. If P3H3 is an LH1 chaperone that is critical for LH1 activity, P3H3 and LH1 null mice should display a similar deficiency in lysyl hydroxylation. To test this hypothesis, we compared the amount and location of hydroxylysine in the triple helical domains of type V and I collagen from P3H3 null, LH1 null, and wild-type mice. The amount of hydroxylysine in type V collagen was reduced in P3H3 null mice, but surprisingly type V collagen from LH1 null mice contained as much hydroxylysine as type V collagen from wild-type mice. In type I collagen, our results indicate that LH1 plays a global enzymatic role in lysyl hydroxylation. P3H3 is also involved in lysyl hydroxylation, particularly at cross-link formation sites, but is not required for all lysyl hydroxylation sites. In summary, our study suggests that LH1 and P3H3 likely have two distinct mechanisms to recognize different collagen types and to distinguish cross-link formation sites from other sites in type I collagen.
- Published
- 2021