A novel pyrroleninone cross-link from bovine dentine

• Published in: Biochimica et biophysica acta Vol. 1381; no. 2; pp. 179 - 190
• Main Authors: Kleter, G.A., Damen, J.J.M., Kettenes-van den Bosch, J.J., Bank, R.A., te Koppele, J.M., Veraart, J.R., ten Cate, J.M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 23.07.1998
• Subjects: Agrotechnological Research Institute; Amino Acids - chemistry; Amino Acids - isolation & purification; Animals; Cattle; Chromatography, Ion Exchange; Collagen; Collagen - chemistry; Cow; Cross-linking amino acid; Cross-Linking Reagents; Dentin - chemistry; Dentine; Electrophoresis, Capillary; Instituut voor Agrotechnologisch Onderzoek; Magnetic Resonance Spectroscopy; Mass Spectrometry; Molecular Structure; Pyrroles - chemistry; Pyrroles - isolation & purification; BAW, n-butanol/acetic acid/water=4/1/1 by volume; Hyl, hydroxylysine; HP, hydroxylysylpyridinoline (pyridinoline); RP, reverse-phase; ( Δ-)DHLNL, (didehydro)dihydroxylysinonorleucine; Cross-linking amino acid; MALDI-TOF, matrix-assisted laser-desorption ionisation time-of-flight; Cow; LP, lysylpyridinoline (deoxypyridinoline); ( Δ-)HLNL, (didehydro)hydroxylysinonorleucine; FAB, fast atom bombardment; ES, electrospray; FMOC, 9-fluorenylmethyloxycarbonyl; SCX, strong cation exchange; Collagen; FMOC-Cl, 9-fluorenylmethyl chloroformate; IP, ion-pair; LAL, lysinoalanine; TOCSY, total correlation spectroscopy; Dentine; PAW, n-propanol/ammonia/water=8/1/11 by volume; CE-DAD, capillary electrophoresis with diode-array detection
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/S0304-4165(98)00027-0

The aim was to identify suspect collagen cross-links in dentine, eluting close to known cross-links in ion-exchange HPLC. Bovine tooth roots as source of dentine were powdered, demineralised, reduced, and acid-hydrolysed. Cross-linking amino acids were isolated from the acid hydrolysate by size exclusion, adsorption, and sequential ion exchange chromatography. In addition to dihydroxylysinonorleucine and hydroxylysylpyridinoline, an unknown cross-link was isolated (V-2). The ultraviolet, mass, and nuclear magnetic resonance spectra support the proposed structure of V-2, a trimeric amino acid with a pyrroleninone nucleus.