Abstract

Lysyl hydroxylase 2 (LH2) is a member of LH family that catalyzes the hydroxylation of lysine (Lys) residues on collagen, and this particular isozyme has been implicated in various diseases. While its function as a telopeptidyl LH is generally accepted, several fundamental questions remain unanswered: 1. Does LH2 catalyze the hydroxylation of all telopeptidyl Lys residues of collagen? 2. Is LH2 involved in the helical Lys hydroxylation? 3. What are the functional consequences when LH2 is completely absent? To answer these questions, we generated LH2-null MC3T3 cells (LH2KO), and extensively characterized the type I collagen phenotypes in comparison with controls. Cross-link analysis demonstrated that the hydroxylysine-aldehyde (Hylald)-derived cross-links were completely absent from LH2KO collagen with concomitant increases in the Lysald-derived cross-links. Mass spectrometric analysis revealed that, in LH2KO type I collagen, telopeptidyl Lys hydroxylation was completely abolished at all sites while helical Lys hydroxylation was slightly diminished in a site-specific manner. Moreover, di-glycosylated Hyl was diminished at the expense of mono-glycosylated Hyl. LH2KO collagen was highly soluble and digestible, fibril diameters were diminished, and mineralization impaired when compared to controls. Together, these data underscore the critical role of LH2-catalyzed collagen modifications in collagen stability, organization and mineralization in MC3T3 cells.

Details

Title
Lysyl hydroxylase 2 mediated collagen post-translational modifications and functional outcomes
Author
Terajima, Masahiko 1 ; Taga, Yuki 2 ; Nakamura, Tomoyuki 3 ; Guo, Hou-Fu 4 ; Kayashima, Yukako 5 ; Maeda-Smithies, Nobuyo 5 ; Parag-Sharma, Kshitij 6 ; Kim, Jeong Seon 4 ; Amelio, Antonio L. 7 ; Mizuno, Kazunori 2 ; Kurie, Jonathan M. 8 ; Yamauchi, Mitsuo 9 

 University of North Carolina at Chapel Hill, Division of Oral and Craniofacial Health Sciences, Adams School of Dentistry, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208) 
 Nippi Research Institute of Biomatrix, Ibaraki, Japan (GRID:grid.10698.36) 
 Kansai Medical University, Department of Pharmacology, Osaka, Japan (GRID:grid.410783.9) (ISNI:0000 0001 2172 5041) 
 University of Kentucky, Department of Molecular and Cellular Biochemistry, Kentucky, USA (GRID:grid.266539.d) (ISNI:0000 0004 1936 8438) 
 University of North Carolina at Chapel Hill, Department of Pathology and Laboratory Medicine, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208) 
 University of North Carolina at Chapel Hill, Graduate Curriculum in Cell Biology & Physiology, Biological & Biomedical Sciences Program, UNC School of Medicine, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208) 
 University of North Carolina at Chapel Hill, Division of Oral and Craniofacial Health Sciences, Adams School of Dentistry, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208); University of North Carolina at Chapel Hill, Department of Cell Biology and Physiology, UNC School of Medicine, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208); University of North Carolina at Chapel Hill, Biomedical Research Imaging Center, UNC School of Medicine, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208); University of North Carolina at Chapel Hill, Lineberger Comprehensive Cancer Center, Cancer Cell Biology Program, UNC School of Medicine, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208) 
 University of Texas MD Anderson Cancer Center, Department of Thoracic/Head and Neck Medical Oncology, Houston, USA (GRID:grid.240145.6) (ISNI:0000 0001 2291 4776) 
 University of North Carolina at Chapel Hill, Division of Oral and Craniofacial Health Sciences, Adams School of Dentistry, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208); University of North Carolina at Chapel Hill, Division of Oral and Craniofacial Health Sciences, Adams School of Dentistry, Chapel Hill, USA (GRID:grid.10698.36) (ISNI:0000000122483208) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2705224033
Copyright
© The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.