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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Exp Eye Res. 2013 Feb 11;109:51–59. doi: 10.1016/j.exer.2013.01.016

Fig. 2. C18 RP-HPLC elution profile of protease-rich fraction of human (20 years) lens extract and MCA-62-70 and LC-MS results.

Fig. 2

MCA-62-70 substrate (MCA-SEVRSDRD[Lys(Dnp)]RR, 50 μg) was incubated with protease-rich human (20 years) lens extract (5 mg) for 5 h at 37 °C. The hydrolyzed peptides were separated from bulk of the proteins by 1:1 isopropanol extraction. The extracted peptides were eluted through C18 RP-HPLC column ( Inline graphic). The elution was monitored using a fluorescence detector with excitation 320 nm and emission 405 nm. A similar sample without incubation served as a control ( Inline graphic). The collected fractions of peaks were analyzed by LC-MS analysis. Hydrolytic cleavages were observed at 62-65 (MCA-SEVR), 62-66 (MCA-SEVRS) and 62-68 (MCA-SEVRSDR) regions, with 62-66 being the predominant.