A single-column chromatographic technique for the analysis of hydroxylysine glycosides and acid hydrolyzates is described. This technique employs a Durrum D-500 amino acid analyzer equipped with a standard 48 cm X 1.8 mm column packed with DC-4A resin. Resolution was achieved with four sodium citrate buffers and four column temperatures. Products of glycoprotein hydrolysis including cysteic acid, methionine sulfoxide, 4-hydroxyproline, alpha-aminobutyric acid, glucosamine, galactosamine, hydroxylysine, tryptophan and the internal standard, norleucine, are resolved. The completely automated procedure takes 185 min per run and can measure amino acid residues in the nanomole range.
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http://dx.doi.org/10.1016/0021-9673(75)85060-6 | DOI Listing |
Int J Biol Macromol
December 2024
Center for Biotechnology, Anna University, Chennai 600 025, India. Electronic address:
Alpha-ketoglutarate (αKG) dependent Lysyl hydroxylase (LH) is a critical enzyme in the post-translational conversion of lysine into hydroxylysine in collagen triple helix and telopeptide regions. Overexpression of LH increases collagen hydroxylation and covalent cross-linkage, causing fibrosis. Currently, no drugs are available to inhibit LH potentially.
View Article and Find Full Text PDFEBioMedicine
January 2025
Department of Public Health Sciences, University of Virginia, School of Medicine, Charlottesville, VA, USA; Department of Family Medicine, University of Virginia, School of Medicine, Charlottesville, VA, USA.
Background: Sudden Infant Death Syndrome (SIDS) is a leading cause of postneonatal mortality. The absence of specific biomarkers of SIDS diagnosis and risk leaves a significant gap in understanding SIDS pathophysiology. Metabolomics offers an avenue to better understand SIDS biology and identifying potential biomarkers.
View Article and Find Full Text PDFFront Bioeng Biotechnol
October 2024
Large Molecule Research, Roche Pharma Research and Early Development (pRED), Roche Innovation Center Munich, Penzberg, Germany.
We present a detailed mass spectrometric analysis of three 2 + 1 T-cell bispecific monoclonal antibodies (TCB mAbs), where an unexpected +15.9950 Da mass shift in tryptic peptides was observed. This modification was attributed to the occurrence of 5R-hydroxylysine (Hyl) using a hybrid LC-MS/MS molecular characterization and CRISPR/Cas9 gene deletion approach.
View Article and Find Full Text PDFChem Sci
October 2024
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota at Twin Cities Minneapolis Minnesota USA
Lysine 5-hydroxylation (5-Hyl) has been well recognized as an essential protein post-translational modification regulating cellular structural stability, RNA alternative splicing and epigenetic gene expression. System-wide enrichment and quantification of 5-Hyl targets have been challenging due to their chemical inert nature and difficulties in differentiating structural isomers in a complex biological sample. Here, we report the development of an efficient chemical proteomic workflow for affinity enrichment and constitutional isomer specific profiling of endogenous 5-Hyl substrates based on highly selective periodate chemistry.
View Article and Find Full Text PDFACS Omega
June 2024
Laboratory of Organic Chemistry, Faculty of Sciences and Environment, Dunarea de Jos University of Galati, Galati 800008, Romania.
Recently, obtaining collagen films using a cross-linking technique has been a successful strategy. The current investigation used six cross-linker extracts (CE) from six different pomegranate varieties' byproducts to make and characterize collagen-tannin films using acid-soluble collagen (SC). The polymeric film has a yellow hue after CE incorporation.
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