Expression and characterization of the thylakoid lumen protease DegP1 from Arabidopsis.

Abstract:

:The Arabidopsis genome contains 14 genes encoding the serine protease DegP. Products of four of these genes are located in the chloroplast: three in the thylakoid lumen and one on the stromal side of the membrane. We expressed the gene encoding DegP1 as a His-tagged fusion protein in Escherichia coli, purified the protein by affinity chromatography, and characterized it biochemically. Size-exclusion chromatography suggested that DegP1 eluted from the column as a mixture of monomers and hexamers. Proteolytic activity was characterized using beta-casein as a model substrate. DegP1 demonstrated concentration-dependent activity, a pH optimum of 6.0 and increasing activity at elevated temperatures. DegP1 was capable of degrading two lumenal proteins, plastocyanin and OE33, suggesting a role as a general-purpose protease in the thylakoid lumen. The results of this work are discussed in the context of the recent elucidation of the structure of the E. coli homolog and the possible physiological role of the protease in the chloroplast lumen.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Chassin Y,Kapri-Pardes E,Sinvany G,Arad T,Adam Z

doi

10.1104/pp.007922

subject

Has Abstract

pub_date

2002-10-01 00:00:00

pages

857-64

issue

2

eissn

0032-0889

issn

1532-2548

journal_volume

130

pub_type

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