Molecular and functional characterization of a cyclophilin with antifungal activity from Chinese cabbage.
An antifungal protein that inhibits the growth of filamentous fungal pathogens was isolated from Chinese cabbage (Brassica campestris L. ssp. pekinensis) by affinity chromatography on Affi-gel blue gel and ion exchange chromatography on CM-Sepharose. The N-terminal amino acid sequence of the protein was highly homologous to that of plant cyclophilins and consequently the protein was denoted as C-CyP. To understand the antifungal activity of C-CyP, we isolated a cDNA encoding its gene from a Chinese cabbage leaf cDNA library. The Chinese cabbage genome bears more than one C-CyP gene copy and C-CyP mRNA is highly expressed in all tissues except the seeds. Recombinant C-CyP catalyzed the cis-trans inter-conversion of the Ala-Pro bond of the substrate, which indicates this protein has peptidyl-prolyl cis-trans isomerase activity. It also inhibited the growth of several fungal pathogens.
Lee
Jung Ro
JR
Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, 660-701, Republic of Korea.
Park
Seong-Cheol
SC
Kim
Jin-Young
JY
Lee
Seung Sik
SS
Park
Yoonkyung
Y
Cheong
Gang-Won
GW
Hahm
Kyung-Soo
KS
Lee
Sang Yeol
SY
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification; D001483:Base Sequence; D020171:Botrytis / Q000187:drug effects; D001937:Brassica / Q000737:chemistry*; D002176:Candida albicans / Q000187:drug effects; D021983:Cyclophilins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D004926:Escherichia coli / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005670:Fusarium / Q000187:drug effects; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000494:pharmacology; D012232:Rhizoctonia / Q000187:drug effects; D016415:Sequence Alignment
D000935:Antifungal Agents; D011994:Recombinant Proteins; D021983:Cyclophilins; D016650:Fluorescein-5-isothiocyanate