Effect Of Wheat Germ Agglutinin Research Articles

EDRF release from canine coronary artery by lectins.

1. The effect of wheat germ agglutinin (WGA), concanavalin A (Con A) and lentil lectin agglutinin (LCA) were investigated on pre-contracted canine coronary artery rings in vitro. 2. In endothelium-intact canine coronary artery, contracted with the thromboxane A2-analogue, U46619, WGA relaxed the tissue in a concentration-dependent manner, with an inhibitory concentration (IC50) of 112 +/- 17 nM (n = 6). In the absence of an endothelium, WGA did not cause any relaxation of the tissue. 3. In endothelium-intact canine coronary artery, contracted with the thromboxane A2-analogue, U46619. LCA relaxed the tissue in a concentration-dependent manner, with an inhibitory concentration (IC50) of 423.1 +/- 41 nM (n = 6). In the absence of an endothelium, LCA produced a 20.1 +/- 1.1% (n = 6) relaxation at the highest concentration tested (3 microM). 4. Concanavalin A (Con A) relaxed canine coronary artery in a partial endothelium-dependent manner with an IC50 of 104 +/- 19 nM on endothelium-intact coronary artery and an IC50 of 1.3 +/- 0.3 microM (n = 6) on endothelium-denuded tissues. 5. The relaxation effects of WGA were attenuated by 1 mM NG-monomethyl L-arginine (L-NMMA) and completely inhibited by haemoglobin (3 microM), methylene blue (10 microM) and LY 83583 (30 microM). Ibuprofen had no effect on WGA-induced relaxation. 6. The relaxant effects of WGA were reversed by addition of 20 mM N-acetyl-D-glucosamine (GlcNAc) and N-acetyl-D-galactosamine (Ga1NAc) but not by alpha-mannose, D-(+)-galactose, and beta-lactose, whereas the endothelin-dependent relaxations to LCA and Con A were unaffected. 7.The endothelium-dependent relaxation induced by the lectins was unaffected by pretreatment of the tissue with 1 microM atropine.8. In the absence of extracellular calcium, WGA was also able to release EDRF suggesting that WGA acts through a second messenger system to release intracellular calcium.9. We suggest that WGA acts as an agonist to release EDRF from endothelial cells possibly by binding to a sugar moiety, specific receptor or adhesion molecules on the endothelial cell surface.

The effect of wheat germ agglutinin on dinitrogen fixation, glutamine synthetase activity and ammonia excretion in Azospirillum brasilense Sp 245

Wheat germ agglutinin was found to increase glutamine synthetase and nitrogenase activities and excretion of N2-fixation product (NH4+) in Azospirillum brasilense Sp 245. Each effect had a similar pattern and correlated well with each other. They were dynamic, had maxima in the middle of the exponential phase of growth, and were due to N-acetyl-d-glucosamine specificity of wheat germ agglutinin, since its preincubation with 10% N-acetyl-d-glucosamine caused the disappearance of the effects. When wheat germ agglutinin was replaced with Ulex europaeus agglutinin II possessing the same carbohydrate binding specificity all the above effects remained, replacing wheat germ agglutinin with concanavalin A with a different sugar specificity, or with bovine serum albumin led to their disappearance.

Microinjected U snRNAs are imported to oocyte nuclei via the nuclear pore complex by three distinguishable targeting pathways.

The inhibitory effects of wheat germ agglutinin and mAb 414 on the nuclear import of all types of U snRNAs indicate that they cross the nuclear envelope through the nuclear pore complex. However, the import of different U snRNAs occurs by kinetically distinct targeting pathways that can be distinguished from one another by the competitive effects of free trimethylguanosine cap dinucleotide (m3GpppG) and P(Lys)-BSA, an efficient synthetic karyophile based on the nuclear localization signal of SV40 large T antigen. The import of U snRNAs that contain 5' m3GpppN caps and are complexed by Sm proteins (U1, U2, U4, and U5) is competed by coinjection with free m3GpppG, indicating a shared transport factor, but not by P(Lys)-BSA. The import of U6 snRNA, which lacks a m3GpppN cap and is not complexed by the Sm proteins, is competed by P(Lys)-BSA but not by free m3GpppG. Thus, by the criterion of kinetic competition, U6 snRNA import is identical to that of the karyophilic proteins P(Lys)-BSA and nucleoplasmin. Uniquely, the import of U3 snRNA, which contains a m3GpppN cap but does not bind Sm proteins is not competed by either free m3GpppG or P(Lys)-BSA. Thus, U3 snRNA appears to be imported by a novel third kinetic pathway.

CMP-N-acetylneuraminic acid hydroxylase activity determines the wheat germ agglutinin-binding phenotype in two mutants of the lymphoma cell line MDAY-D2

The dominant glycosylation mutants of MDAY-D2 mouse lymphoma cells, designated class 2 (D33W25 and D34W25) were selected for their resistance to the toxic effects of wheat germ agglutinin (WGA) and shown to express elevated levels of Neu5Gc. In accordance with this, the activity of CMP-Neu5Ac hydroxylase was found to be substantially higher in the mutant cells. The hydroxylase in the D33W25 mutant cells exhibited kinetic properties identical to those of the same enzyme from mouse liver. Growth rate experiments in vivo and in vitro, where the mutant cells grew more slowly at low cell densities in serum-free medium and also formed slower growing tumours in syngeneic mice, indicate that CMP-Neu5Ac hydroxylase expression may be associated with altered growth of the mutant cells.

Evidence of Glycosylated Sites on the Endothelin-1 Receptor in Swiss 3T3 Cells

The effects of incubation of intact cells with six different lectins on the specific binding of [125I]endothelin-1 (ET-1) were determined in Swiss 3T3 fibroblasts. ET-1 binding was unaffected by pretreatment of cells for 1 h at 37 degrees C with concanavalin A, soybean agglutinin, Ulex europaeus agglutinin I, peanut agglutinin, or Galanthus nivalis agglutinin. However, preincubation of cells with 300 micrograms/ml of wheat germ agglutinin resulted in a 70% decrease in specific binding of ET-1 to cell-surface receptors. The inhibitory effects of wheat germ agglutinin were diminished by brief incubation of lectin-treated cells with 100 mM N-acetylglucosamine, a monosaccharide specifically recognized by wheat germ agglutinin. Neither glucose nor mannose had any effect on wheat germ agglutinin-mediated inhibition of the specific binding of ET-1. These results suggest that the ET-1 receptor on 3T3 cells is a glycoprotein that contains one or more N-acetylglucosamine residues at or near the ligand binding site.

Sugar-Specific Inhibitory Effects of Wheat Germ Agglutinin and Phytohemagglutinin-E4 on Histamine Release Induced by Basic Secretagogues from Rat Peritoneal Mast Cells and Their Possible Action Sites

The histamine release induced by compound 48/80, bradykinin or polyethylenimine with a molecular weight of 600 (PEI6) was inhibited by wheat germ agglutinin (WGA) and phytohemagglutinin E-subunits (PHA-E4), and the inhibition was specifically reversed by N-acetyl glucosamine and N-acetyl galactosamine, respectively. Concanavalin A (Con A) and phytohemagglutinin L-subunits (PHA-L4) did not inhibit the histamine release induced by compound 48/80, bradykinin or PEI6. The histamine release induced by substance P was also inhibited sugar-specifically by WGA and PHA-E4. The binding sites for compound 48/80, bradykinin, PEI6 and substance P, therefore, seemed to especially overlap each other. These binding sites were found to be glycoproteins having affinities to WGA and PHA-E4, but not to Con A and PHA-L4. The binding of WGA and PHA-E4 to the glycoproteins resulted in inhibition of the interaction between the basic secretagogues including bradykinin and substance P and their binding sites on the mast cells. The bindings of five lectins to mast cell glycoproteins were examined by lectin-blotting. Several glycoproteins, which had specific affinities to WGA and PHA-E4, but not to Con A and PHA-L4 were detected. We assumed that the binding sites for basic secretagogues which are coupled with histamine-releasing mechanisms exist among these glycoproteins. A 41-kDa protein (alpha-subunit of pertussis toxin-sensitive G protein) was not detected by WGA, suggesting that the binding sites for the basic secretagogues were not G proteins.

Wheat germ agglutinin inhibits thrombin-induced rises in cytosolic free calcium and prostacyclin synthesis by human umbilical vein endothelial cells.

To characterize the endothelial cell surface membrane glycoproteins that mediate thrombin stimulation of PGI2 synthesis by human umbilical vein endothelial cells (HUVEC), HUVEC were stimulated with thrombin in the presence or absence of different lectins. Of the lectins tested, only wheat germ agglutinin (WGA) inhibited thrombin-induced rises in cytosolic free calcium [( Ca2+]i), measured using Quin 2-loaded HUVEC and PGI2 production measured by radioimmunoassay. However, WGA by itself had no influence on baseline [Ca2+]i or PGI2 production and did not inhibit histamine-induced rises in [Ca2+]i. The inhibition of thrombin-induced rises in [Ca2+]i and PGI2 production by WGA was dose dependent, with half-maximal inhibition occurring at 2 micrograms/ml. WGA also inhibited thrombin-induced release of 3H-arachidonic acid. These effects of WGA were reversed by N-acetyl-glucosamine (GlcNAc) and N-acetyl-neuraminic acid, which bind specifically to WGA, but not by unrelated sugars. Succinylated WGA (succ-WGA), a chemically modified derivative of WGA that binds to GlcNAc but, unlike native WGA, not to sialoglycoproteins, did not inhibit thrombin-induced rises in [Ca2+]i and PGI2 production. These results suggest that thrombin induces rises in [Ca2+]i and PGI2 production by interacting with an endothelial surface membrane sialoglycoprotein.

Effect of wheat germ agglutinin on T lymphocyte activation.

Wheat germ agglutinin (WGA) is low mitogenic or nonmitogenic for human T lymphocytes and inhibits phytohemagglutinin (PHA)-induced mitotic response of the lymphocytes. In this study, the effect of WGA was analyzed in terms of interleukin 2 (IL2) production, expression of IL2 receptor, and IL2 responsiveness of the T lymphocytes. WGA as well as PHA could induce IL2 mRNA and IL2 production and also elevate cytoplasmic free Ca2+ concentration. The IL2 production was reduced by inhibitors of calmodulin and protein kinase C. The IL2 receptor (Tac) expression was induced at about 20% of the lymphocytes by WGA and the expression induced by PHA was not blocked by the addition of WGA. The lymphocytes precultured with WGA for 3 days could proliferate by the addition of IL2 after removal of WGA. The IL2-dependent proliferation of PHA-blasts was blocked by the addition of WGA. These results indicate that WGA inhibits T lymphocyte proliferation by inhibiting the responsiveness of the lymphocytes to IL2 but not by interfering with IL2 production and IL2 receptor expression.

Direct effects of wheat germ agglutinin on inositol phosphate formation and cytosolic-free calcium level in intestine 407 cells.

The interaction between dietary lectins, especially wheat germ agglutinin (WGA), and intestinal cells has been implicated in the pathogenesis of celiac disease. The present study was undertaken to investigate the immediate effects following such an interaction. Direct WGA-stimulation of Intestine 407 cells leads to an immediate rise in the cytosolic-free calcium concentration. The major part of this lectin-induced rise is due to an influx of calcium across the plasma membrane into the cytosol. However, WGA-exposure also results in an immediate mobilization of calcium from intracellular stores, most likely mediated via the simultaneous increase of inositol trisphosphate formation in these cells. The transduction mechanism described for WGA in these intestinal cells is not very sensitive towards pertussis toxin, indicating that if a G-protein is involved, it differs from those of most other systems. The suggested role for WGA in changing the functional and structural properties of intestinal cells might involve increases of inositol phosphate and cytosolic-free calcium levels.

Depressed lymphocyte responses to wheat germ agglutinin and other mitogens in treated coeliac disease patients

Abstract This report investigates the possibility that inappropriate lymphocyte responses to wheat germ agglutinin (WGA), a mitogen found in wheat which influences proliferation of and immunoglobulin (Ig) synthesis by peripheral blood mononuclear cells (PBMC), may account for some of the immunological dysfunction noted in coeliac disease. The PBMC were prepared from healthy controls and 16 patients with coeliac disease who had received a gluten‐free diet for periods of 1 month to over 20 years. There was no difference in the proliferative responses of PBMC from the patients and normal subjects to optimal mitogenic concentrations of phytohaemagglutinin (PHA) or Concanavalin A (Con A), but the responses of the cells of the patients to pokeweed mitogen (PWM) and to a suboptimal concentration of Con A were reduced. The proliferative response induced by WGA in PBMC from treated coeliac patients was significantly depressed compared with controls. The synthesis of Ig by PWM‐stimulated PBMC, and the Con A induced suppression of this synthesis, was the same in cultures of PBMC from the patients or the controls. There was no difference in the effect of WGA on Ig synthesis between the patients and controls. It is concluded that there is no difference in the effect of WGA on Ig synthesis by PBMC from coeliac disease patients or controls, but that lymphocytes from coeliac disease patients proliferate poorly in response to WGA.

Cytotoxicity of lectins on rat intestinal mucosa enhanced by neuraminidase.

The lectins wheat germ agglutinin (WGA) and Concanavalin A (ConA) were perfused into an isolated small intestinal segment alone or after prior perfusion with neuraminidase for a 10 day period in the rat. Intestinal morphometry, intraepithelial Lymphocyte (IEL) and round cell content as well as digestive capacity was measured in the loop and in the adjacent segments. Both lectins induce a mucosal transformation in all segments but ConA is more effective than WGA. Pre-incubation with neuraminidase enhances the action of ConA throughout, whereas only a partial enhancement of the effect of WGA is observed. The mucosal transformation after long term perfusion with the lectins resembles the hyperregenerative adaptation of the mucosa due to gluten in coeliac disease and may thus serve as an animal model for this disease.

Effect of wheat germ agglutinin on the interleukin pathway of human T lymphocyte activation.

Wheat germ agglutinin (WGA) inhibits proliferation of human peripheral blood mononuclear cells (PBMC) induced by mitogens and antigens. We investigated the mechanism by which WGA inhibits PHA-induced human lymphocyte proliferation with regard to the interleukin pathway. Our data revealed that although PBMC-proliferation was markedly suppressed by WGA, levels of IL 2 activity in WGA-inhibited cultures were not reduced, but instead were increased, suggesting failure to utilize IL 2. Furthermore, the addition of exogenous IL 2 failed to overcome the suppression. Consistent with these observations, culturing PBMC with PHA plus WGA markedly decreased the number of high-affinity IL 2 receptor per cell, as determined by binding of purified [3H]IL 2, relative to cultures containing PHA alone. WGA immobilized on support beads bound detergent-solubilized IL 2 receptors from PHA-activated T cells, but did not bind human IL 2. However, WGA did not competitively block the binding of [3H]IL 2 to PHA-induced lymphoblasts. These results suggest that WGA inhibits lymphocyte proliferation by binding to and decreasing the number of high-affinity IL 2 receptors displayed on T cells, without impairing IL 2 production.

2,2,6,6-Tetramethyl-4-Amino[d17]Piperidine-15N-1-OXYL as a Spin Label For Cell Surface Carbohydrates

Abstract The first use of perdeuterated, 15N-Tempamine spin label to selectively monitor cell surface sialic acid of human erythrocyte membranes is reported. The molecular reasons for significantly increased sensitivity and significantly decreased scatter in the data over that of protonated, 14N-Tempamine are discussed. These advantages are illustrated by monitoring the effects of wheat-germ agglutinin on the motion of terminal sialic acid of membrane glycoconjugates.

Effect of altered oligosaccharide structure on the cell surface number, distribution and turnover of the high molecular weight acidic glycoproteins of CHO cells.

The influence of altered carbohydrate structure on the surface number, distribution and turnover of plasma membrane glycoproteins has been studied in Chinese hamster ovary (CHO) cells by comparing three lines that are resistant to the cytotoxic effects of wheat germ agglutinin (WGA) with parental CHO cells. The glycoproteins investigated were members of a group of high molecular weight acidic glycoproteins (HMWAG). On parental cells these represent the major surface components that become labelled by lactoperoxidase-catalysed iodination. They are the only plasma membrane glycoproteins that bind WGA. The mutant lines also possess iodinatable surface polypeptides of high molecular weight, but these were less acidic and electrophoretically less diffuse than those from parental cells. These polypeptides in general did not bind [125I]WGA when two-dimensional polyacrylamide gels were overlaid with iodinated lectin. Mutant cells treated with fluorescein-conjugated WGA showed low surface fluorescence. However, the nuclear envelope and a small region in the perinuclear zone fluoresced strongly. Together, these results confirm that the surface glycoproteins of mutant cells had altered carbohydrate structure. Mouse antiserum prepared against the HMWAG, however, bound equally effectively to the mutant lines as to the parental lines. Indirect immunofluorescence experiments showed that the HMWAG had a fairly uniform distribution over the surface, and that internalization induced by second antibody occurred at a similar rate and in a similar manner in all lines, including the mutants. Electron microscopic observations using immunoperoxidase procedures confirmed the similarities in glycoprotein distribution on mutant and parental cells. Two mouse monoclonal antibodies raised against the HMWAG also revealed no difference in the number or topography of surface glycoproteins. Finally, the half-lives of several HMWAG in a parental and a mutant line (15B) maintained on low-serum medium were compared by means of a 125I/131I double-label technique. Half-lives of HMWAG from the former averaged 12 h and from the latter 11 h. It is concluded that the lack of complex termini on oligosaccharides of this particular group of CHO plasma membrane glycoproteins has no effect on their number, distribution or turnover.

Wheat germ agglutinin decreases expression of the T8 antigen on human peripheral mononuclear cells.

We examined dynamics of expression of the human T-suppressor specific antigen, T8, following interaction of peripheral lymphocytes with wheat germ agglutinin (WGA). Cells were incubated at 37 degrees C with or without WGA (15 micrograms/ml) for 18 hrs, washed sequentially with N-acetylglucosamine (to remove bound WGA) and plain medium, then analyzed by flow cytometry for binding of lectins and monoclonal antibodies OKT8(T-suppressor specific) and OKT3 (pan-T specific). WGA pretreatment induced an overall 65% reduction in WGA binding and concomitant 30% reduction in percentage of T8+ cells. Furthermore, residual T8+ cells showed 50% reduction in T8 expression. Taking into account reductions in both percentages of T8+ cells and also antigen densities, WGA reduced T8 expression by greater than 60% overall. By contrast, binding of OKT3 and the lectins, concanavalin A (con A) and Ricinus communis agglutinin (RCA-I), was unaffected by WGA. The decreased T8 expression could not be explained by residual cell bound WGA and was fully reversible within 48 hours of removal of cells from WGA-containing medium. Therefore, WGA caused downregulation of T8 antigen expression. The effect of WGA was time- and concentration-dependent. Downregulation did not occur at 4 degrees C nor in the presence of azide, thereby demonstrating a requirement for cellular metabolism. The data suggest that WGA may bind to the T8 antigen, and they provide the possibility that similar downregulation of T8 by WGA may underlie certain of the in vitro immunoregulatory effects of this lectin.

Inhibition of human lymphocyte activation by wheat germ agglutinin: A model for saccharide-specific suppressor factors

The suppressive effect of wheat germ agglutinin (WGA) on lectin-stimulated blastogenesis and immunoglobulin production was studied. Addition of WGA at 10 μg/ml inhibited phytohemagglutinin (PHA)-, concanavalin-A (Con-A)-, and pokeweed mitogen (PWM)-induced mitogenic responses by 70–80%. PWM-driven immunoglobulin synthesis was suppressed by 45% with WGA. The inhibitory effects of WGA were not due to cell death or to interference with lectin binding at the cell surface. Inhibition was dependent on the presence of WGA in the cell culture during the first 24 hr of mitogen exposure and was observed in cultures of both monocytedepleted peripheral blood mononuclear cells as well as T-cell-enriched populations. WGA-induced inhibition of blastogenesis was blocked by the addition of N-acetylglucosamine (GluNAc) which prevents WGA binding to the cell surface. WGA was found to mimic the suppressive effect of a soluble immune suppressor supernatant (SISS) derived from Con-A-activated mononuclear cell cultures. PHA responses were inhibited by 80 and 95% with SISS and WGA, respectively. The inhibition by both WGA and SISS was totally reversed with addition of GluNAc. Furthermore, WGA and SISS demonstrated competition for the same cell surface receptor site. WGA may therefore be useful as an in vitro model of a saccharide-specific, biologically relevant, soluble mediator for the investigation of mechanisms of immunologic suppression.

Influence of lectins, hexoses, and neuraminidase on the association of purified elementary bodies of Chlamydia trachomatis UW-31 with HeLa cells

Using highly purified elementary bodies of Chlamydia trachomatis UW-31 (serotype K), we found that HeLa 229 monolayer cultures bound more 32P-labeled chlamydiae after pretreatment with the lectin wheat germ agglutinin. The lectin, on the other hand, inhibited competitively when chlamydial association was assayed in the presence of polycations. The two effects of wheat germ agglutinin were abolished when N-acetylneuraminic acid (NeuNAc)- or N-acetylglucosamine (GlcNAc)-preincubated wheat germ agglutinin was used. Brief exposure of HeLa cells to neuraminidase abolished the ability to bind the elementary bodies, whether or not polycations were present. Furthermore, at 5 degrees C but not at 37 degrees C, NeuNAc, GlcNAc and N-acetylgalactosamine inhibited chlamydial association only in the absence of the polycation DEAE-dextran. The results suggest that NeuNAc residues on the plasma membrane are the principal, but not the only, receptors for this strain of C. trachomatis.

Change in state of nerve growth factor receptor. Modulation of receptor affinity by wheat germ agglutinin.

The binding of 125I-labeled nerve growth factor (NGF) to human melanoma cell (A875) membranes, detergent-soluble membrane extracts, and membrane extracts reconstituted into phospholipid vesicles was significantly increased when binding was carried out in the presence of wheat germ agglutinin (WGA). In the absence of WGA, all 125I-NGF binding was rapidly eliminated by trypsin treatment or rapidly dissociated in the presence of a high concentration of unlabeled NGF. However, in the presence of WGA, up to 75% of 125I-NGF bound was resistant to trypsin digestion and was only slowly dissociated by a high concentration of unlabeled NGF. The effects of WGA can be blocked or reversed by N-acetylglucosamine. Both WGA and NGF rapidly associate with soluble extracts and reconstituted vesicles and, at the concentrations used here, reach binding equilibrium within 2 min. The conversion to slowly dissociating, trypsin-resistant binding, however, was not complete for at least 10 min. Both WGA and NGF are required for maximum accumulation of trypsin-resistant, slowly dissociating binding. The order of addition of NGF and WGA has no effect on the rate of conversion of NGF-receptor, and the conversion occurs after both NGF and WGA are present. The amount of conversion is dependent on the incubation temperature, and significantly greater conversion occurs at 37 than at 0 degrees C. The generation of the trypsin-resistant, slowly dissociating state of NGF-receptor is consistent with a time- and temperature-dependent conformational change in NGF-receptor which occurs after interaction of both NGF and WGA with the receptor or closely associated structures.

Wheat germ agglutinin specifically inhibits formyl peptide-induced polymorphonuclear leukocyte chemotaxis.

Evidence that surface membrane glycoproteins of polymorphonuclear leukocytes (PMN) are involved in stimulus-response coupling prompted us to examine effects on these cells of various plant lectins. We have found that wheat germ agglutinin (WGA) (1.0 microgram/ml) completely, specifically, and irreversibly inhibits directed migration (chemotaxis) of human PMN toward the synthetic peptide, N-formylmethionyl-leucyl-phenylalanine (FMLP) (0.1 to 100 nM). This effect of WGA was not shared by subagglutinating concentrations of either concanavalin A or Bandeirea simplicifolia lectin. In contrast to its effects on FMLP-induced chemotaxis, WGA did not influence other FMLP-induced PMN responses (i.e., selective discharge of lysosomal enzymes from cytochalasin B-treated cells, generation of superoxide anion radical(s). WGA also did not influence PMN chemotactic responses to either the complement-derived peptide, C5a, or the lipoxygenase product, leukotriene B4. Inhibition of FMLP-induced chemotaxis by WGA was not reversed by washing WGA-treated cells, but was reversed (and prevented) by N-acetyl-D-glucosamine (not by N-acetyl-D-galactosamine or mannosamine). WGA did not affect either orientation or stimulated random motility of PMN, and did not interfere with specific binding to PMN of (3H)-FMLP. A derivative of WGA with 10-fold less agglutinating activity for human erythrocytes was prepared by treating the native lectin with cyanogen bromide and formic acid. The derivative also inhibited FMLP-induced PMN chemotaxis specifically and selectively. These data suggest that WGA specifically inhibits FMLP-induced PMN chemotaxis by attaching to N-acetyl-D-glucosamine residues at a locus on the PMN plasma membrane that is distinct from the binding site of the FMLP receptor.

Effect of wheat germ agglutinin on the viscoelastic properties of erythrocyte membrane.

The elasticity and viscosity of the human erythrocyte membrane were measured as a function of the concentration of wheat germ agglutinin (WGA) in a suspending solution containing 1 mg/ml albumin, approximately 5 X 10(5) cells/ml and between 0.0 and 0.2 microgram/ml WGA. Membrane elasticity was characterized by the elastic shear modulus, which provided a measure of the resistance of the membrane to constant-area elastic deformations that occurred in the membrane plane. The elastic shear modulus was determined by aspirating a portion of the membrane into a micropipette and measuring the extension of the membrane into the pipette as a function of the suction pressure. The results indicated no significant change in shear modulus for concentrations of WGA between 0.0 and 0.2 microgram/ml. Membrane viscosity was characterized by the coefficient of surface viscosity, which, in effect, was a measure of the membrane's resistance to rates of deformation. This coefficient was determined from the time required for an erythrocyte to recover its undeformed shape after it had been elongated by the application of an equal and opposite force applied at diametrically opposite points on the erythrocyte rim. The value for the coefficient of surface viscosity was found to increase by a factor of almost three when the WGA concentration was increased from 0.0 to 0.2 microgram/ml. These results indicated that, in the presence of albumin, WGA can increase membrane dissipation (viscosity) without altering the structural rigidity (elasticity) of the membrane.