Failure to find Ca2(+)-dependent proteinase (calpain) activity in a plant species, Elodea densa.
Five and nine-tenth kg of Elodea densa (Anacharis), a common aquarium plant, was extracted, and the extract was subjected to column chromatographic procedures that successfully purify the two Ca2(+)-dependent proteinases (calpains) and their protein inhibitor (calpastatin) from a variety of animal tissues. Although these procedures purified a protein having 55- and 16-kDa polypeptides, neither this protein nor any of the other chromatographic fractions contained detectable proteinase or calpastatin activity. Moreover, the purified 55- and 16-kDa polypeptides did not react on immunoblots with polyclonal antibodies that were monospecific for the calpains or calpastatin. We conclude that Elodea densa contains no calpain nor calpastatin at the level of 4 micrograms per g plant protein (1 part per 250,000), which was the sensitivity of our assay.
Wolfe
F H
FH
Departments of Animal Sciences, University of Arizona, Tucson 85721.
Szpacenko
A
A
McGee
K
K
Goll
D E
DE
D000199:Actins / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002154:Calpain / Q000378:metabolism*; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D007382:Intermediate Filaments / Q000187:drug effects; D010944:Plants / Q000201:enzymology*
D000199:Actins; D002135:Calcium-Binding Proteins; C033668:calpastatin; D002154:Calpain; D002118:Calcium