Lysyl hydroxylase 2 mediated collagen post-translational modifications and functional outcomes

Masahiko Terajima, Yuki Taga, Tomoyuki Nakamura, Hou Fu Guo, Yukako Kayashima, Nobuyo Maeda-Smithies, Kshitij Parag-Sharma, Jeong Seon Kim, Antonio L. Amelio, Kazunori Mizuno, Jonathan M. Kurie, Mitsuo Yamauchi

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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.

Original languageEnglish
Article number14256
JournalScientific Reports
Issue number1
StatePublished - Dec 2022

Bibliographical note

Funding Information:
This work was supported by NIH R01CA251067 and SPORE P50CA070907 to JMK and MY, Basic Science Fund from Nippi to YT, JSPS KAKENHI Grant Number JP19H03439 to TN, NIH R01 HL049277 to NMS, Developmental Research Program Grant from the Yale Head and Neck SPORE NIDCR P50-DE030707 to ALA, NIH R00 CA225633 to HG. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Publisher Copyright:
© 2022, The Author(s).

ASJC Scopus subject areas

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