Collagen prolyl 4-hydroxylase isoenzymes I and II have sequence specificity towards different X-Pro-Gly triplets

• DOI: 10.1016/j.matbio.2023.12.001
• Publication Date: 2023-12-09T16:30:43Z
• Authors: Antti M. Salo; Pekka Rappu; M．Kristian Koski; Emma Karjalainen; Valerio Izzi; Kati Drushinin; Ilkka Miinalainen; Jarmo Käpylä; Jyrki Heino; Johanna Myllyharju
• Keywords: Hydroxyproline; Hydroxylation; Imino acid; Collagen helix; Type I collagen; Hydroxylysine

Collagen biosynthesis requires several co- and post-translational modifications of lysine proline residues to form structurally functionally competent collagen molecules. Formation 4-hydroxyproline (4Hyp) in Y-position prolines the repetitive -X-Y-Gly- sequences provides thermal stability for triple-helical 4Hyp formation is catalyzed by a prolyl 4-hydroxylase (C-P4H) family consisting three isoenzymes. Here we identify specific roles two main C-P4H isoenzymes hydroxylation detailed analysis type I IV collagens derived from cell tissue samples. Loss C-P4H-I results underhydroxylation where affected are not uniformly distributed, but mainly present sites adjacent X-position amino acid has positively charged or polar uncharged side chain. In contrast, loss C-P4H-II triplets occupied negatively glutamate aspartate. Hydroxylation these was found be important as alone resulted reduced melting temperature altered assembly fibrils basement membrane. The observed isoenzyme differences substrate specificity were explained selective binding active site resulting distinct Km Vmax values. Furthermore, our clearly show that selection dependent on type, determinant -X-Pro-Gly- triplet. Although data shows necessity both II normal 4-hydroxylation function collagens, mRNA expression with various procollagens was, surprisingly, tightly coordinated, suggesting additional levels control. conclusion, this study molecular level explanation need multiple generate molecules capable assemble into intact extracellular matrix structures.