Production of an antibody specific for the propeptide of wheat germ agglutinin

Journal Article · Sun Oct 01 04:00:00 EDT 1989 · Plant Physiology; (USA)

DOI:https://doi.org/10.1104/pp.91.2.473· OSTI ID:5947916

Smith, J J; Raikhel, N V ^[1]

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1. Michigan State Univ., East Lansing (USA)

Wheat germ agglutinin (WGA) is synthesized as a proprotein with a glycosylated, 15 amino acid, carboxyl-terminal propeptide. This glycopeptide is cleaved from pro-WGA to produce the mature lectin during the transport of WGA to the protein bodies/vacuoles. To study the posttranslational modification of WGA, it would be useful to be able to differentiate between pro-WGA and mature WGA. Therefore, a peptide corresponding to the propeptide of WGA was synthesized (WGA-B 172-186), and an antiserum was raised in rabbits (anti-WGA-B 172-186). Anti-WGA-B 172-186 reacted with pure WGA-B 172-186 and pro-WGA in ELISA. Anti-WGA-B 172-186 was also specific for and readily differentiated between pro-WGA and mature WGA on Western blots. This provided an assay to monitor pro-WGA on Western blots before and after endo-{beta}-N-acetylglucosaminidase H digestion. Using this assay, direct evidence was obtained that the oligosaccharide of pro-WGA is of the high-mannose type.

• OSTI ID: 5947916
• Journal Information: Plant Physiology; (USA), Journal Name: Plant Physiology; (USA) Vol. 91:2; ISSN 0032-0889; ISSN PLPHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANTIGENS
BIOASSAY
CARBOHYDRATES
CEREALS
GRASS
HEXOSES
IMMUNE SERUMS
IMMUNOASSAY
LECTINS
LILIOPSIDA
MAGNOLIOPHYTA
MANNOSE
MONOSACCHARIDES
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
PEPTIDES
PLANTS
PROTEINS
SACCHARIDES
WHEAT