Two bean cell wall proteins more abundant during water deficit are high in proline and interact with a plasma membrane protein (original) (raw)
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Planta, 2007
Plant cell walls undergo dynamic changes in response to diVerent environmental stress conditions. In response to water deWcit, two related proline-rich glycoproteins, called p33 and p36, accumulate in the soluble fraction of the cell walls in Phaseolus vulgaris (Covarrubias et al. in Plant Physiol 107:1119-1128. In this work, we show that p33 and p36 are able to form a 240 kDa oligomer, which is found in the cell wall soluble fraction. We present evidence indicating that the highest accumulation of these proteins in response to water deWcit occurs in the growing regions of common bean seedlings, particularly in the phloem tissues. These proteins were detected in P. vulgaris cell suspension cultures, where the p33/p36 ratio was higher under hyperosmotic conditions than in bean seedlings subjected to the same treatment. The results support a role for these proteins during the plant cell response to changes in its water status, and suggest that cell wall modiWcations are induced in active growing cells of common bean in response to water limitation.
Cell-Wall Proteins Induced by Water Deficit in Bean (Phaseolus vulgaris L.) Seedlings
Plant physiology, 1995
In the last few years, much attention has been given to the role of proteins that accumulate during water deficit. In this work, we analyzed the electrophoretic patterns of basic protein extracts, enriched for a number of cell-wall proteins, from bean (Phaseolus vulgaris L.) seedlings and 21-d-old plants subjected to water deficit. Three major basic proteins accumulated in bean seedlings exposed to low water potentials, with apparent molecular masses of 36, 33, and 22 kD, which we refer to as p36, p33, and p22, respectively. Leaves and roots of 21-d-old plants grown under low-water-availability conditions accumulated only p36 and p33 proteins. In 21-d-old plants subjected to a fast rate of water loss, both p33 and p36 accumulated to approximately the same levels, whereas if the plants were subjected to a gradual loss of water, p33 accumulated to higher levels. Both p36 and p33 were glycosylated and were found in the cell-wall fraction. In contrast, p22 was not glycosylated and was f...
A plant surface protein sharing structural properties with animal integrins
1998
Using a polyclonal antibody (P23) generated against the human platelet integrin AIIbβ3 and a FITCconjugate secondary antibody, fluorescence is observed at the surface of protoplasts isolated from Arabidopsis thaliana and Rubus fruticosus. Arabidopsis thaliana cells grown in suspension culture containing P23 and glycylarginylglycylaspartylserine (GRGDS), a synthetic peptide containing the RGD sequence found in many extracellular matrix adhesive proteins demonstrated aberrant cell wall/plasma membrane interactions and organization. When glycoproteins from these plants, purified on a concanavalin A Sepharose 4B, were subjected to SDS/PAGE and Western blotting, under reduced and non-reduced conditions, immunoblots probed with P23 revealed bands in both species. A shift in electrophoretic mobility is observed to different apparent molecular mass when no reducing agent is present. When purified by immunoaffinity chromatography on anti-AIIbβ3 Sepharose or Sepharose linked to the synthetic ...
Isolation of Plant Cell Wall Proteins
Methods in Molecular Biology™, 2008
The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins; (iii) the presence of proteins interacting in many different ways with the polysaccharide matrix require different procedures to elute them from the cell wall. Three categories of CWP are distinguished: labile proteins that have little or no interactions with cell wall components, weakly bound proteins extractable with salts, and strongly bound proteins. Two alternative protocols are decribed for cell wall proteomics: (i) non-destructive techniques allowing the extraction of labile or weakly bound CWP without damaging the plasma membrane; (ii) destructive techniques to isolate cell walls from which weakly or strongly bound CWP can be extracted. These protocols give very low levels of contamination by 2 intracellular proteins. Their application should lead to a realistic view of the cell wall proteome at least for labile and weakly bound CWP extractable by salts.
Plasma membrane protein trafficking in plant-microbe interactions: a plant cell point of view
Frontiers in Plant Science, 2014
In order to ensure their physiological and cellular functions, plasma membrane (PM) proteins must be properly conveyed from their site of synthesis, i.e., the endoplasmic reticulum, to their final destination, the PM, through the secretory pathway. PM protein homeostasis also relies on recycling and/or degradation, two processes that are initiated by endocytosis. Vesicular membrane trafficking events to and from the PM have been shown to be altered when plant cells are exposed to mutualistic or pathogenic microbes. In this review, we will describe the fine-tune regulation of such alterations, and their consequence in PM protein activity. We will consider the formation of intracellular perimicrobial compartments, the PM protein trafficking machinery of the host, and the delivery or retrieval of signaling and transport proteins such as pattern-recognition receptors, producers of reactive oxygen species, and sugar transporters. FIGURE 1 | Perimicrobial membranes in three types of plant-microbe interactions. Location of proteins specifically targeted to or excluded from the perimicrobial membranes are indicated. Endomembrane compartments or vesicles involved are illustrated. Small GTPases and exocyst subunits are not represented. AGP, arabinogalactan protein; CW, cell wall; EHM, extrahaustorial membrane; E. matrix, extrahaustorial matrix; ER, endoplasmic reticulum; FLOT, flotillin; LYK3, lysM-family receptor-like kinase from Medicago; MVB, multivesicular body; N, fixed-nitrogen; Nod53b, nodule-specific 53-kDa protein from soybean; NPSN, novel plant SNARE, PAM, periarbuscular membrane; PAS, periarbuscular space; PBM, peribacteroid membrane; PIP, plasma membrane intrinsic protein (aquaporin); PM, plasma membrane; PT, phosphate transporter; REM, remorin; ROR2, required for mlo-specified resistance2 syntaxin; ROS, reactive oxygen species; RPW8.2, resistance powdery mildew8.2; SYMREM, symbiotic remorin1; SYP, syntaxin of plant; TGN, trans-Golgi network; VAMP, vesicle-associated membrane protein; VYP, vapyrin.
Large scale characterization of plant plasma membrane proteins
Biochimie, 1999
After a brief review of the strategies used to date to identify systematically plasma membrane (PM) proteins, emphasis was given to the proteomic approach of PM proteins from the model plant Arabidopsis thaliana. Comparative analysis of two-dimensional gels from PM and cytosolic fractions was used to assess the cellular origin of proteins found in PM fraction. The classification obtained was confirmed by protein sequencing that showed, in addition, that most analyzed proteins were peripheral proteins. A large proportion of these appeared to correspond to PM-constitutive proteins that were present in the PM from different plant organs, but were not uniquely located at the PM depending on the organ. In addition, the presence of organ-specific sets of PM-specific proteins was also demonstrated. Additional procedures were developed to identify integral PM proteins. The combined use of PM washes with alkaline carbonate buffer or Triton X-100/KBr, and of a new detergent to solubilize protein, resulted in improved recovery of hydrophobic proteins on gels. Results are discussed in terms of construction of comprehensive proteomes for PM and other membranes and organelles. © Société française de biochimie et biologie moléculaire / Elsevier, Paris Arabidopsis thaliana / plasma membrane / proteome / two-dimensional gel electrophoresis * Correspondence and reprints Biochimie 81 (1999) 655−661 © Société française de biochimie et biologie moléculaire / Elsevier, Paris
Integrin-Like Proteins are Localized to Plasma Membrane Fractions, not Plastids, in Arabidopsis
Plant and Cell Physiology, 1999
Integrins are a large family of integral membrane proteins that function in signal transduction in animal systems. These proteins are conserved in vertebrates, invertebrates, and fungi. Evidence from previous research suggests that integrin-Iike proteins may be present in plants as well, and that these proteins may function in signal transduction during gravitropism. In past studies, researchers have used monoclonal and polyclonal antibodies to localize fi 1 integrin-like proteins in plants. However, there is a disparity between data collected from these studies, especially since molecular weights obtained from these investigations range from 55-120 kDa for integrin-like proteins. To date, a complete investigation which employs all three basic immunolabeling procedures, immunoblotting, immunofluorescence microscopy, and immunogold labeling, in addition to extensive fractionation and exhaustive controls, has been lacking. In this paper, we demonstrate that use of a polyclonal antibody against the cytoplasmic domain of avian /?i-integrin can produce potential artifacts in immunolocalization studies. However, these problems can be eliminated through use of starchless mutants or proper specimen preparation prior to electrophoresis. We also show that this antibody, when applied within the described parameters and with careful controls, identifies a large (100 kDa) integrin-like protein that is localized to plasma membrane fractions in Arabidopsis.
Secondary Cell-Wall-Specific Glycoprotein(s) from French Bean Hypocotyls
Plant Physiology, 1995
Specific labeling of secondary cell walls of tracheary elements and of xylary and phloem fibers has been observed when wheat germ agglutinin (WCA) and anti-WCA antibodies were used during ultrastructural studies of French bean (Phaseolus vulgaris L.) hypocotyls. In this report we demonstrate that at least part of this labeling is due to the presence of secondary cell-wall-specific glycoproteins. Three major nove1 glycoproteins with relative molecular weights of 55,000, 86,000, and 90,000, purified by means of WCA-Sepharose affinity chromatography, have been characterized. Their amino acid composition indicates that they are not the members of known classes of structural cell-wall proteins, since they contain no hydroxyproline, a lower level of glycine than seen in glycine-rich proteins, and very little proline. N-terminal sequences of all three proteins show no significant homology with other proteins. Antibodies were raised against electrophoretically pure 90-kD glycoprotein. These were used to localize this protein in secondary cell walls of xylem tracheary elements and in xylary and phloem fibers, i.e. in the same compartments where labeling with WCA has been observed. To our knowledge this is one of the first biochemical and ultrastructural demonstrations of secondary cellwall-specific gl ycoproteins.
A family of Arabidopsis plasma membrane receptors presenting animal β-integrin domains
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1999
A cDNA clone, AtELP1 (Arabidopsis thaliana EGF receptor-like protein) was isolated from an Arabidopsis cDNA library with an oligonucleotide probe corresponding to a highly conserved region of animal L-integrins. The cloning of this cDNA was previously reported and it has been proposed that AtELP might be a receptor involved in intracellular trafficking. In the present work, using two specific independent sets of anti-peptide antibodies, we show that AtELP1 is mainly located in the plasma membrane, supporting another function for this protein. Structural studies, using methods for secondary structure prediction, indicated the presence of cysteine-rich domains specific to L-integrins. Database searches revealed that AtELP1 is a member of a multigenic family composed of at least six members in A. thaliana. Northern blot analysis of AtELP1, 2b and 3 was performed on mRNA extracted from cells cultured in normal and stressed conditions, and from several organs and plants submitted to biotic or abiotic stresses. All the genes are expressed at different levels in the same conditions, but preferentially in roots, fruits and leaves in response to water deficit. ß 0167-4838 / 99 / $^see front matter ß 1999 Elsevier Science B.V. All rights reserved. PII: S 0 0 0 5 -2 7 2 8 ( 9 9 ) 0 0 0 8 7 -0 * Corresponding