Chemical synthesis, expression and mutagenesis of a gene encoding beta-cryptogein, an elicitin produced by Phytophthora cryptogea.
Elicitins are 10 kDa holoproteins secreted by Phytophthora fungi, that elicit an incompatible hypersensitive reaction, leading to resistance against fungal and bacterial plant pathogens. Comparison of primary sequences of alpha-elicitins and beta-elicitins indicated several potential necrotic activity-determining residues. All of the highly necrotic beta-elicitins have a hydrophilic residue (usually lysine) at position 13, whereas in the less necrotic alpha-elicitins this residue is replaced by a valine. Here, we report the synthesis and expression of a gene encoding a highly necrotic elicitin, beta-cryptogein, and we show that the substitution of Lys-13 of this recombinant protein by a valine leads to a drastic alteration to the necrotic activity of the recombinant protein.
O'Donohue
M J
MJ
Laboratoire d'Etude des Protéines INRA, Jony-en-Josas, France.
Gousseau
H
H
Huet
J C
JC
Tepfer
D
D
Pernollet
J C
JC
D020418:Algal Proteins; D000595:Amino Acid Sequence; D001483:Base Sequence; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D015870:Gene Expression; D017343:Genes, Plant / Q000235:genetics*; D005813:Genes, Synthetic / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010838:Phytophthora / Q000235:genetics*; D010947:Plants, Toxic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D014026:Nicotiana / Q000187:drug effects
D020418:Algal Proteins; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; C061032:cryptogein protein, Phytophthora cryptogea