Zsuzsanna Darula, Maxwell C. McCabe, Alex Barrett, Lauren R. Schmitt, Mark D. Maslanka, Anthony J Saviola, Joseph Orgel, Alma Burlingame, Claudia A. Staab-Weijnitz, Kurt Stenmark, Valerie Weaver, Robert J. Chalkley, Kirk C. Hansen
Hungarian Centre of Excellence for Molecular Medicine. HUN-REN Biological Research Centre. University of Colorado Denver, Anschutz Medical Campus. Illinois Institute of Technology. University of California San Francisco.
Hungary and United States
Matrix Biology Plus
Matrix Biol Plus 2025;
DOI: 10.1016/j.mbplus.2025.100179
Abstract
Collagen cross-links mediated by the lysyl oxidase and lysyl hydroxylase families of enzymes significantly contribute to the biomechanical strength and rigidity of tissues, influencing cell signaling and the downstream cell phenotype. In the clinic, the proteolytically liberated N-terminal cross-linked peptide of collagen I (NTX) is used as a biomarker of bone and connective tissue turnover, which is altered in several disease processes. Despite the clinical utility of these collagen breakdown products, the majority of the cross-linked peptide species have not been identified in proteomic datasets. Here, we evaluate several parameters for the preparation and identification of these peptides from the collagen I-rich Achilles tendon. Our refined approach, which involves chemical digestion for protein solubilization coupled with mass spectrometry, enables the identification of NTX cross-links in a range of modification states. We then applied a spectral library approach to identify differences in collagen cross-links in bovine pulmonary hypertension. The presented method offers unique opportunities to understand extracellular matrix remodeling events in development, aging, wound healing, and fibrotic disease that modulate collagen architecture through lysyl hydroxylase and lysyl oxidase enzymes.
Category
Animal Models of Pulmonary Vascular Disease and Therapy
Class III. Pulmonary Hypertension Associated with Alveolar Hypoxia
Vascular Cell Biology and Mechanisms of Pulmonary Vascular Disease
Potential Biomarkers Associated with Pulmonary Vascular Disease
Age Focus: No Age-Related Focus
Fresh or Filed Publication: Fresh (PHresh). Less than 1-2 years since publication
Article Access
Free PDF File or Full Text Article Available Through PubMed or DOI: Yes