Abstract
Ubiquitous type m-calpain and lens specific Lp82 calpain were separated and partially purified from fetal bovine lens and the enzymatic characteristics were compared. Lens m-calpain required 200 μM calcium for 1/2 maximal activity, while Lp82 required 30 μM. Both types of calpains were inhibited by 0.1 mM E64, and 5 mM iodoacetamide, but not by 1 mM phenylmethylsulfonyl fluoride. Lp82 was insensitive to 1 μM calpastatin peptide while m-calpain was effectively inhibited. In the presence of calcium, m-calpain lost most of its activity within 2 hr, while Lp82 was continually active for 18 hr. Both calpains cleaved the natural substrates βA3 and αB crystallins in a similar manner. However, incubation of αA crystallin with m-calpain removed ten amino acid residues from its C-terminus, while incubation with Lp82 removed only five residues. The latter truncation product of αA was also found in vivo. These data suggested that Lp82 may have a more important role than m-calpain in modification of crystallins during lens maturation.
Original language | English (US) |
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Pages (from-to) | 625-637 |
Number of pages | 13 |
Journal | Experimental Eye Research |
Volume | 73 |
Issue number | 5 |
DOIs | |
State | Published - 2001 |
Keywords
- Alpha crystallin
- Beta crystallin
- Bovine lens
- Lp82: m-calpain
- Mass spectrometry
- Maturation
- Proteolysis
ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
- Cellular and Molecular Neuroscience