UNLABELLED: To elucidate the roles of LH2b in collagen cross-linking, MC3T3-E1 cell clones expressing higher (S) or lower (AS) levels of LH2b were established. Compared with controls, the collagen cross-linking pattern was shifted toward hydroxylysine-aldehyde (S clones)- or lysine-aldehyde (AS clones)-derived pathways. The data indicate that LH2b directs collagen cross-linking pathways through its action on telopeptidyl lysine residues. INTRODUCTION: Lysine (Lys) hydroxylation is a post-translational modification of collagen critical for cross-linking and glycosylation. Currently, three isoforms of lysyl hydroxylase (LH) have been identified, but their specific functions are still not well defined. Recently, we proposed that LH2 might modulate collagen cross-linking pattern through its action on Lys residues located in the telopeptide domains of collagen. MATERIALS AND METHODS: To directly test this hypothesis, several MC3T3-E1 cell-derived clones expressing higher (sense [S]) or lower (antisense [AS]) levels of LH2b, the predominant form of LH2 in this cell line, were established and cultured for 2 weeks, and collagen cross-links and precursor aldehydes in the matrices were analyzed. RESULTS: In S clones tested, the ratio of dihydroxylysinonorleucine (DHLNL) to hydroxylysinonorleucine (HLNL) was significantly higher than the average of controls (76% and 140% increase, respectively), and the level of pyridinoline (Pyr) was elevated (100% and 150% increase, respectively). In contrast, when MC3T3-E1 cells were transfected with a LH2b antisense construct (AS clones), the DHLNL/HLNL ratios were significantly lower than that of controls (56% and 73% decrease, respectively), and Pyr was not detected. Furthermore, significant amounts of an aldol-derived cross-link, dehydrohistidinohydroxymerodesmosine, were produced ( approximately 0.3 mol/mol of collagen) in AS clones. CONCLUSIONS: The data clearly show a critical role of LH2b in determining collagen cross-linking pathways, most likely through its action on telopeptidyl Lys residues.
UNLABELLED: To elucidate the roles of LH2b in collagen cross-linking, MC3T3-E1 cell clones expressing higher (S) or lower (AS) levels of LH2b were established. Compared with controls, the collagen cross-linking pattern was shifted toward hydroxylysine-aldehyde (S clones)- or lysine-aldehyde (AS clones)-derived pathways. The data indicate that LH2b directs collagen cross-linking pathways through its action on telopeptidyl lysine residues. INTRODUCTION:Lysine (Lys) hydroxylation is a post-translational modification of collagen critical for cross-linking and glycosylation. Currently, three isoforms of lysyl hydroxylase (LH) have been identified, but their specific functions are still not well defined. Recently, we proposed that LH2 might modulate collagen cross-linking pattern through its action on Lys residues located in the telopeptide domains of collagen. MATERIALS AND METHODS: To directly test this hypothesis, several MC3T3-E1 cell-derived clones expressing higher (sense [S]) or lower (antisense [AS]) levels of LH2b, the predominant form of LH2 in this cell line, were established and cultured for 2 weeks, and collagen cross-links and precursor aldehydes in the matrices were analyzed. RESULTS: In S clones tested, the ratio of dihydroxylysinonorleucine (DHLNL) to hydroxylysinonorleucine (HLNL) was significantly higher than the average of controls (76% and 140% increase, respectively), and the level of pyridinoline (Pyr) was elevated (100% and 150% increase, respectively). In contrast, when MC3T3-E1 cells were transfected with a LH2b antisense construct (AS clones), the DHLNL/HLNL ratios were significantly lower than that of controls (56% and 73% decrease, respectively), and Pyr was not detected. Furthermore, significant amounts of an aldol-derived cross-link, dehydrohistidinohydroxymerodesmosine, were produced ( approximately 0.3 mol/mol of collagen) in AS clones. CONCLUSIONS: The data clearly show a critical role of LH2b in determining collagen cross-linking pathways, most likely through its action on telopeptidyl Lys residues.
Entities:
Keywords:
NASA Discipline Cell Biology; Non-NASA Center
Authors: Yulong Chen; Masahiko Terajima; Yanan Yang; Li Sun; Young-Ho Ahn; Daniela Pankova; Daniel S Puperi; Takeshi Watanabe; Min P Kim; Shanda H Blackmon; Jaime Rodriguez; Hui Liu; Carmen Behrens; Ignacio I Wistuba; Rosalba Minelli; Kenneth L Scott; Johannah Sanchez-Adams; Farshid Guilak; Debananda Pati; Nishan Thilaganathan; Alan R Burns; Chad J Creighton; Elisabeth D Martinez; Tomasz Zal; K Jane Grande-Allen; Mitsuo Yamauchi; Jonathan M Kurie Journal: J Clin Invest Date: 2015-02-09 Impact factor: 14.808
Authors: Johanna A Knipper; Sebastian Willenborg; Jürgen Brinckmann; Wilhelm Bloch; Tobias Maaß; Raimund Wagener; Thomas Krieg; Tara Sutherland; Ariel Munitz; Marc E Rothenberg; Anja Niehoff; Rebecca Richardson; Matthias Hammerschmidt; Judith E Allen; Sabine A Eming Journal: Immunity Date: 2015-10-20 Impact factor: 31.745
Authors: Hou-Fu Guo; Eun Jeong Cho; Ashwini K Devkota; Yulong Chen; William Russell; George N Phillips; Mitsuo Yamauchi; Kevin N Dalby; Jonathan M Kurie Journal: Arch Biochem Biophys Date: 2017-02-20 Impact factor: 4.013
Authors: Maryna Perepelyuk; Masahiko Terajima; Andrew Y Wang; Penelope C Georges; Paul A Janmey; Mitsuo Yamauchi; Rebecca G Wells Journal: Am J Physiol Gastrointest Liver Physiol Date: 2013-01-17 Impact factor: 4.052