Purchase this article with an account.
Paul N. Bishop, David McLeod, Anthony Reardon; Effects of Hyaluronan Lyase, Hyaluronidase, and Chondroitin ABC Lyase on Mammalian Vitreous Gel. Invest. Ophthalmol. Vis. Sci. 1999;40(10):2173-2178.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
purpose. To determine the effects of enzymes on mammalian vitreous gel and to
thus infer the structural roles of hyaluronan and chondroitin sulfate
in the gel.
methods. The wet weights of bovine vitreous gels were compared before and after
incubation with Streptomyces hyaluronan lyase, chondroitin
ABC lyase, testicular hyaluronidase, or buffer alone. The extent of
hyaluronan depolymerization was determined by chromatography and that
of chondroitin sulfate depolymerization by western blot analysis.
results. After digestion with Streptomyces hyaluronan lyase (30
U/gel), the gel wet weight was the same as that of controls (incubated
with buffer alone) despite 94% of the hyaluronan having been
depolymerized; when digested with 100 U/gel, the gel wet weight
decreased (to 57% of original wet weight versus 86% for controls, P = < 0.001) and hyaluronan was completely
depolymerized. Chondroitin ABC lyase digestion (0.2 U/gel) resulted in
a slight reduction in gel wet weight (90% versus 96%, P = < 0.001) and depolymerization of 88% of the
hyaluronan; the presence of fully digested chondroitin sulfate chains
was established. Digestions with 100 and 500 U/gel of testicular
hyaluronidase resulted in a decrease (P = < 0.001,
both cases) in gel wet weight (53% versus 82%, 100 U/gel; 57% versus
86%, 500 U/gel) with 75% and 97% hyaluronan depolymerization,
conclusions. Depolymerization of all vitreous hyaluronan and of chondroitin sulfate
resulted in gel wet weight reduction but not gel destruction. Digestion
with 30 U/gel of Streptomyces hyaluronan lyase revealed
a small pool (6%) of relatively enzyme-resistant hyaluronan that
specifically contributed toward maintaining gel wet
This PDF is available to Subscribers Only