Identification, isolation, and N-terminal sequencing of style glycoproteins associated with self-incompatibility in Nicotiana alata.
S-Gene-associated glycoproteins (S-glycoproteins) from styles of Nicotiana alata, identified by non-equilibrium two-dimensional electrophoresis, were purified by cation exchange fast protein liquid chromatography with yields of 0.5 to 8 micrograms of protein per style, depending on the S-genotype of the plant. The method relies on the highly basic nature of the S-glycoproteins. The elution profiles of the different S-glycoproteins from the fast protein liquid chromatography column were characteristic of each S-glycoprotein, and could be used to establish the S-genotype of plants in outbreeding populations. In all cases, the S-genotype predicted from the style protein profile corresponded to that predicted from DNA gel blot analysis using S-allele-specific DNA probes and to that established by conventional breeding tests. Amino-terminal sequences of five purified S-glycoproteins showed a high degree of homology with the previously published sequences of N. alata and Lycopersicon esculentum S-glycoproteins.
Jahnen
W
W
Plant Cell Biology Research Centre, School of Botany, University of Melbourne, Parkville, Victoria, Australia.
Batterham
M P
MP
Clarke
A E
AE
Moritz
R L
RL
Simpson
R J
RJ
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006023:Glycoproteins / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification*; D010947:Plants, Toxic; D014026:Nicotiana / Q000737:chemistry* / Q000235:genetics
D006023:Glycoproteins; D010940:Plant Proteins; C076697:self-incompatibility glycoprotein, Nicotiana