hydroxylysine and Lathyrism

The hydroxylation of lysine in embryonic chick long bone and mandibular collagen was found to be approximately 3-fold greater than that of the collagens of adult animals. In contrast, no significant difference was found in extent of lysine hydroxylation of the collagens of frontal bones of embryos and postnatal animals. Both histochemical and biochemical evidence established that full thickness diaphyseal bone samples contained cartilage and, consequently, type II collagen which undoubtedly contributed to the higher hydroxylysine contents of young postnatal animals reported previously. DEAE ion exchange chromatography of the alpha 1(I) chains of lathyritic long bone and mandibular collagens isolated by carboxymethyl-cellulose ion exchange chromatography showed considerable heterogeneity, whereas the alpha 1(I) chains obtained from lathyritic frontal bone collagen did not. Three fractions of alpha 1(I) chains of long bones and mandibular collagen were isolated which differed significantly in their hydroxylysine contents. The relative proportion of the three peaks changed as a function of embryonic age and maturation: more of the alpha 1(I) chains with the highest hydroxylysine content was present in the collagen synthesized earliest during embryonic development. This is consistent with results which demonstrated that the collagens synthesized earliest during embryonic and postnatal development had the highest hydroxylysine contents.

Topics: Aging; Animals; Bone and Bones; Bone Development; Chick Embryo; Chickens; Collagen; Hydroxylysine; Lathyrism; Lysine; Skull

alpha- and beta-Chains were isolated by sequential ion-exchange and gel-filtration chromatography of guanidinium chloride-soluble dentine collagen obtained from Tris/NaCl-extracted EDTA-demineralized lathyritic-rat incisors. The alpha-chains were identified as alpha 1 I and alpha 2 by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and amino acid analysis of the intact chains and their CNBr peptides. The dentine alpha-chains exhibited higher lysine hydroxylation and phosphate content, but lower hydroxylysine glycosylation, than alpha-chains from skin. Increased lysine hydroxylation was observed in the helical sequences. The alpha 1 I/alpha 2 ratio was approx. 3:1, and was presumably due to the presence of (alpha 1 I)3 molecules along with (alpha 1 I)2 alpha 2 molecules as shown recently for neutral-salt-soluble dentine collagen [Wohllebe & Carmichael (1978) Eur. J. Biochem. 92, 183--188]. In the borohydride-reduced beta 11- and beta 12-chains from guanidinium chloride-soluble dentine collagen, the reduced cross-links hydroxylysinohydroxynorleucine and hydroxylysinonorleucine were present. A higher proportion of hydroxylysinonorleucine in the reduced beta 12-chain probably reflects differences in extent of hydroxylation of specific lysine residues of the alpha 1 I- and alpha 2-chains.

Topics: Amino Acids; Animals; Chemical Phenomena; Chemistry; Chromatography, Ion Exchange; Collagen; Cross-Linking Reagents; Dentin; Electrophoresis, Polyacrylamide Gel; Glycosides; Guanidines; Hydroxylysine; Incisor; Lathyrism; Peptide Fragments; Phosphates; Rats

The affinity of a bone collagen alpha1 chain for hydroxyapatite has been compared to a similarly isolated skin component from the same animals. The chain from bone exhibits a higher affinity for the mineral. This enhanced affinity appears to be related to its primary structure, specifically its hydroxylysine moieties.

Topics: Age Factors; Aminopropionitrile; Animals; Bone and Bones; Chromatography; Collagen; Hydroxyapatites; Hydroxylysine; Lathyrism; Male; Methods; Rats; Skin; Structure-Activity Relationship

Glucosylation of galactosylhydroxylysyl residues in various collagen polypeptide chains and in small peptides prepared from collagen was studied in vitro using collagen glucosyltransferase purified about 200 to 500-fold from extract prepared from chick embryos. When various denatured polypeptide or peptide chains were compared as substrates for the enzyme, no significant differences were found between citrate-soluble collagens from normal or lathyritic rats and isolated alpha1 and alpha2 chains. In contrast, gelatinized insoluble calf skin collagen, and peptides prepared from collagen and having an average molecular weight of about 500 were clearly less effective substrates as judged from their Km and V values. A marked difference was found between native and heat-denatured citrate-soluble collagen in that no synthesis of glucosylgalactosylhydroxylysine was observed with the native collagen when the reaction was studied at 30 degrees C with different times, enzyme concentrations, and substrate concentrations. When the reaction was studied as a function of temperature, little glucosylation of native collagen was observed below 37 degrees C, but there was a sharp transition in the rate of glucosylation of native collagen at temperatures above 37 degrees C, similar to that observable in the melting curve of collagen. The data suggest that triple-helical conformation of collagen prevents that glucosylation of galactosylhydroxylysyl residues.

Topics: Animals; Cattle; Chick Embryo; Citrates; Collagen; Galactose; Glucosyltransferases; Hot Temperature; Hydroxylysine; Kinetics; Lathyrism; Peptide Fragments; Protein Conformation; Protein Denaturation; Rats; Temperature

Topics: Acetates; Amino Acids; Animals; Chickens; Circular Dichroism; Collagen; Electrophoresis, Disc; Hot Temperature; Hydroxylysine; Hydroxyproline; Lathyrism; Microscopy, Electron; Pepsin A; Protein Conformation; Protein Denaturation; Rats; Skin; Species Specificity; Tail; Tendons; Urodela

Topics: Aminopropionitrile; Animals; Ascorbic Acid; Borohydrides; Carbon Isotopes; Cell Line; Collagen; Connective Tissue; Fibroblasts; Glycosaminoglycans; Hydroxylation; Hydroxylysine; Isoniazid; Lathyrism; Lysine; Mice; Oxidation-Reduction; Proline; Protein Biosynthesis; Semicarbazides; Solubility; Time Factors; Tritium

Topics: Amino Acids; Aminopropionitrile; Animals; Cartilage; Chemical Precipitation; Chickens; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Collagen; Dialysis; Hydroxylysine; Lathyrism; Macromolecular Substances; Osmolar Concentration; Protein Conformation; Sodium Chloride

Topics: Aminopropionitrile; Animals; Bone and Bones; Chick Embryo; Chromatography, Gel; Chromatography, Ion Exchange; Collagen; Cyanogen Bromide; Hydroxylysine; Lathyrism; Mixed Function Oxygenases; Peptides; Proline; Protein Denaturation; Pyridines; Rats; Skin; Skull; Tritium; Urea