Synthesis of cross-links in the helical domain of collagen using pyridoxal 5-phosphate and copper or iron
AssigneeThe Regents of the University of California
InventorRobert C. Siegel
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Abstract
Aldehyde cross-link intermediates and cross-links are generated in the central helical portion of collagen by incubating collagen with pyridoxal-5-phosphate and either cupric copper ion or ferrous iron ion. The cross-links are chemically similar to natural cross-links found in the non-helical regions and are directly between amino-acid moieties naturally occuring in the central helical portion of collagen. Cross-linking and utilization of aldehyde intermediates occurs when the product is reincubated after pyridoxal is removed. Alternatively maintaining the product at body temperatures will promote cross-linking. The cross-linked collagen product has increased resistance to enzyme degradation.
Claims18
An in vitro method for increasing the resistance to degradation of a proteolytic enzyme treated and purified collagen product comprising incubating said proteolytic enzyme treated collagen product with pyridoxal 5-phosphate and an ionic metal selected from the group consisting of copper and iron.
An in vitro method for producing native cross-linking sites on a proteolytic enzyme treated and purified collagen product comprising incubating said collagen product with pyridoxal-5-phosphate and metal ion selected from the group consisting of copper and iron.
An in vitro method for producing cross-links in a proteolytic enzyme treated and purified collagen product comprising incubating said collagen product with pyridoxal-5-phosphate and an ionic metal selected from the group consisting of copper and iron, thereafter removing the pyridoxal-5-phosphate and ionic metal from the incubated collagen product, and then reincubating the collagen product alone until cross-links are formed.
An in vitro method for producing cross-linking sites in a product consisting essentially of the central helical portions of collagen comprising incubating said collagen product with pyrodoxal-5-phosphate and metal ion selected from the group consisting of cupric copper and ferrous iron, and continuing the incubation until at least some of the lysyl and hydroxylysyl moieties in the collagen are converted into the aldehyde functional form.
A collagen product consisting essentially of the central helical portion of native collagen and having cross-links of the native type produced by incubating reaction mixture consisting essentially of said collagen with pyridoxal-5-phosphate and either cupric copper ion or ferrous iron ion.
A collagen product consisting essentially of the central helical portion of native collagen and having cross-linking sites produced by incubating reaction mixture consisting essentially of said collagen with pyridoxal-5-phosphate and either cupric copper ion or ferrous iron ion.
Dependencies are inferred from the claim text.