Mutation in the RGD motif decreases the esterase activity of Xcc_est.
Five truncated constructs of Xcc_est GDSL esterase from Xanthomonas campestris were heterologously expressed and purified. The truncated constructs with a RGD motif had higher specific activities than those without the motif. The specific activity of wild-type Xcc_est was 32.5 +/- 2.7 U/mg, while the RGD mutant was 12.5 +/- 4.9 U/mg. Moreover, we expressed mature forms of the Xcc_est protein and the RGD mutant as inclusion bodies and, after refolding, there was no significant difference between the two constructs in specific activity. These results suggest that the RGD motif affects the esterase-domain folding in vivo during the translocation process.
Wang
Jianjun
J
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 100101 Beijing, People's Republic of China.
Cao
Yanping
Y
Zheng
Guojun
G
D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004950:Esterases / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D002479:Inclusion Bodies; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014970:Xanthines / Q000378:metabolism*; D016959:Xanthomonas campestris / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D014970:Xanthines; C054469:8-(4-carboxymethyloxy)phenyl-1,3-dipropylxanthine; D004950:Esterases