Expression and localization of plant protein disulfide isomerase

A cDNA clone encoding a putative protein disulfide isomerase (PDI, EC 5.3.4.1) from alfalfa (Medicago sativa L.) was expressed in Escherichia coli cells, and an antiserum was raised against the expressed PDI-active protein. The antiserum recognized a protein of approximately 60 kD in extracts from a...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 1993-11, Vol.103 (3), p.719-726
Main Authors: Shorrosh, B.S, Subramaniam, J, Schubert, K.R, Dixon, R.A
Format: Article
Language:English
Subjects:
ADN
Alfalfa
Analytical, structural and metabolic biochemistry
Antiserum
Biological and medical sciences
Cell walls
CELLULE
CELULAS
CHAMBRE DE CROISSANCE
CLONE
CLONES
CONTROL AMBIENTAL
CONTROLE DU MILIEU
Cultured cells
Disulfides
Enzymes
Enzymes and enzyme inhibitors
ESCHERICHIA COLI
FITOTRONES
Fundamental and applied biological sciences. Psychology
Gels
GLYCINE MAX
IMMUNSERUM
ISOMERASAS
ISOMERASE
Isomerases
MEDICAGO SATIVA
Metabolism
Metabolism and Enzymology
NICOTIANA TABACUM
Plant physiology and development
Plant roots
Plants
PROTEINAS
PROTEINE
Proteins
Soybeans
SUERO INMUNE
SULFURE
SULFUROS
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Description
Summary:A cDNA clone encoding a putative protein disulfide isomerase (PDI, EC 5.3.4.1) from alfalfa (Medicago sativa L.) was expressed in Escherichia coli cells, and an antiserum was raised against the expressed PDI-active protein. The antiserum recognized a protein of approximately 60 kD in extracts from alfalfa, soybean, and tobacco roots and stems. Levels of this protein remained relatively constant on exposure of alfalfa cell suspension cultures to the protein glycosylation inhibitor tunicamycin, whereas a slightly lower molecular mass form, also detected by the antiserum, was induced by this treatment. A lower molecular mass form of PDI was also observed in roots of alfalfa seedlings during the first 5 weeks after germination. PDI levels increased in developing soybean seeds up to 17 d after fertilization and then declined. Tissue print immunoblots revealed highest levels of PDI protein in the cambial tissues of soybean stems and petioles and in epidermal, subepidermal, cortical, and pith tissues of stems of alfalfa and tobacco. Immunogold electron microscopy confirmed the localization of PDI to the endoplasmic reticulum in soybean root nodules
ISSN:0032-0889
1532-2548
DOI:10.1104/pp.103.3.719