Identification and characterization of DegP, a serine protease associated with the luminal side of the thylakoid membrane.
The proteases involved in proteolytic degradation in the thylakoid lumen are largely unknown. Western analysis with an antibody against the Escherichia coli periplasmic serine protease DegP suggested that pea chloroplasts contain a homologue of this protease. This homologue was peripherally bound to the luminal side of the thylakoid membrane and could only be removed by a combination of high salt and non-ionic detergent. Its level increased almost 2-fold in pea seedlings exposed to elevated temperature for 4 h, suggesting this protease's role in the chloroplast's heat response. Isolated thylakoid membranes containing the chloroplastic homologue of DegP degraded beta-casein, an in vitro substrate of the bacterial protease. This activity was partially inhibited by a serine protease inhibitor, suggesting that at least part of the casein-degrading activity in the thylakoid membrane is attributable to DegP. The existence of chloroplastic DegP was further supported by isolating a full-length Arabidopsis cDNA (designated AtDegP) encoding a protein that is 37% identical and 60% similar to the E. coli protease. The amino terminus of the deduced amino acid sequence contained a bipartite transit peptide, typical of proteins targeted to the thylakoid lumen, and the mature portion of the protein contained the highly conserved serine protease catalytic triad His-Asp-Ser. The possible physiological roles of chloroplastic DegP protease are discussed.
Itzhaki
H
H
Department of Agricultural Botany, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot 76100, Israel.
Naveh
L
L
Lindahl
M
M
Cook
M
M
Adam
Z
Z
D000595:Amino Acid Sequence; D001483:Base Sequence; D002364:Caseins / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology*; D018076:DNA, Complementary; D006360:Heat-Shock Proteins; D006358:Hot Temperature; D006868:Hydrolysis; D007425:Intracellular Membranes / Q000201:enzymology*; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology; D033903:Periplasmic Proteins; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
D002364:Caseins; D018076:DNA, Complementary; D006360:Heat-Shock Proteins; D033903:Periplasmic Proteins; C061682:DegP protease; D012697:Serine Endopeptidases