Purification and characterization of an antifungal protein, C-FKBP, from Chinese cabbage.
An antifungal protein was isolated from Chinese cabbage (Brassica campestris L. ssp. pekinensis) by buffer-soluble extraction and two chromatographic procedures. The results of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that the isolated Chinese cabbage protein was identical to human FK506-binding protein (FKBP). A cDNA encoding FKBP was isolated from a Chinese cabbage leaf cDNA library and named C-FKBP. The open reading frame of the gene encoded a 154-amino acid polypeptide. The amino acid sequence of C-FKBP exhibits striking degrees of identity with the corresponding mouse (61%), human (60%), and yeast (56%) proteins. Genomic Southern blot analyses using the full-length C-FKBP cDNA probe revealed a multigene family in the Chinese cabbage genome. The C-FKBP mRNA was highly expressed in vegetative tissues. We also analyzed the antifungal and peptidyl-prolyl cis-trans isomerase activity of recombinant C-FKBP protein expressed in Escherichia coli. This protein inhibited pathogenic fungal strains, including Candida albicans, Botrytis cinerea, Rhizoctonia solani, and Trichoderma viride, whereas it exhibited no activity against E. coli and Staphylococcus aureus. These results suggest that recombinant C-FKBP is an excellent candidate as a lead compound for the development of antifungal agents.
Park
Seong-Cheol
SC
Environmental Biotechnology National Core Research Center and Division of Applied Life Sciences, Gyeongsang National University, Jinju, Korea.
Lee
Jung Ro
JR
Shin
Sun-Oh
SO
Jung
Ji Hyun
JH
Lee
Young Mee
YM
Son
Hyosuk
H
Park
Yoonkyung
Y
Lee
Sang Yeol
SY
Hahm
Kyung-Soo
KS
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000737:chemistry*; D003001:Cloning, Molecular; D005658:Fungi / Q000187:drug effects; D005659:Fungicides, Industrial / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D015870:Gene Expression; D008969:Molecular Sequence Data; D022021:Tacrolimus Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*
D005659:Fungicides, Industrial; D022021:Tacrolimus Binding Proteins