Quantitation of ultraviolet light-absorbing fractions of the cornea.
This study attempts to measure the quantitative contribution of major chemical fractions of the whole bovine cornea to ultraviolet (UV) absorption between 240 and 300 nm, with special attention to the biologically significant range of 290-300 nm. The cornea was divided into water-insoluble, nonprotein small water-soluble, water-soluble protein, and lipid-soluble fractions. The insoluble fraction (largely collagen) was solubilized by enzymatic digestion. Solutions of individual fractions equivalent to a constant mass of fresh cornea were scanned for absorption from 240 to 300 nm. The sum of the absorbances of the individual fractions closely approximated the absorbance of whole corneas at all wavelengths examined. The extinction coefficients of the lipid and water-soluble fractions were several times greater than that of the insoluble fraction throughout the studied spectrum. Yet, because of its large mass (75% of cornea dry weight), the insoluble fraction accounted for 40-50% of UV absorbance between 240 and 280 nm. However, in the range of 290-300 nm, the water-soluble plus lipid-soluble fractions accounted for 60-65% of the total absorption, with the water-soluble proteins alone accounting for 40-45% of the total. The soluble proteins comprised only approximately 17% of the cornea's dry weight. The special contribution of the water-soluble proteins to absorption was attributed to their relatively high tryptophan content (approximately 1.6% by weight). A 54-kDa protein, identified by others as tryptophan-rich, tumor-associated aldehyde dehydrogenase, accounted for approximately 30% of the total soluble protein mass.(ABSTRACT TRUNCATED AT 250 WORDS)
Mitchell
J
J
Department of Biochemistry, Kirksville College of Osteopathic Medicine, MO 63501, USA.
Cenedella
R J
RJ
D000042:Absorption; D000818:Animals; D002417:Cattle; D003315:Cornea / Q000737:chemistry / Q000528:radiation effects*; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000528:radiation effects*; D008055:Lipids / Q000032:analysis / Q000302:isolation & purification / Q000528:radiation effects; D009696:Nucleic Acids / Q000032:analysis / Q000302:isolation & purification / Q000528:radiation effects; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet; D014466:Ultraviolet Rays
D005136:Eye Proteins; D008055:Lipids; D009696:Nucleic Acids