Purification, immunological characterization and cDNA cloning of a 47 kDa glycoprotein secreted by carrot suspension cells.
A 47 kDa glycoprotein, termed EP4, was purified from carrot cell suspension culture medium. An antiserum raised against EP4 also recognized a protein of 45 kDa that was ionically bound to the cell wall. EP4 was detected in culture media from both embryogenic and non-embryogenic cell lines and was found to be secreted by a specific subset of non-embryogenic cells. Secretion of the 47 kDa glycoprotein by embryogenic cells was not evident. The 45 kDa cell wall-bound EP4 protein was specific for non-embryogenic cells and was shown by immunolocalization to occur in the walls of clustered cells, with the highest levels in the walls separating adjacent cells. In seedlings, EP4 proteins were mainly found in roots. EP4 cDNA was cloned by screening a cDNA library with an oligonucleotide derived from an EP4 peptide sequence. The EP4 cDNA sequence was found to be 55% homologous to ENOD8, an early nodulin gene from alfalfa.
van Engelen
F A
FA
Department of Molecular Biology, Agricultural University Wageningen, The Netherlands.
de Jong
A J
AJ
Meijer
E A
EA
Kuil
C W
CW
Meyboom
J K
JK
Dirkse
W G
WG
Booij
H
H
Hartog
M V
MV
Vandekerckhove
J
J
de Vries
S C
SC
D000595:Amino Acid Sequence; D001483:Base Sequence; D002473:Cell Wall / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018552:Daucus carota / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D015723:Gene Library; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D007106:Immune Sera; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D010940:Plant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D012639:Seeds; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; C093935:EP4 glycoprotein, Daucus carota; D006023:Glycoproteins; D007106:Immune Sera; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D010940:Plant Proteins