Glycobiology Advance Access originally published online on June 29, 2006
Glycobiology 2006 16(10):981-990; doi:10.1093/glycob/cwl019
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Use of lectins for probing differentiated human embryonic stem cells for carbohydrates
2 Department of Biological Chemistry and 3 Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI 48109
1 To whom correspondence should be addressed; e-mail: igoldste{at}umich.edu
Received on April 12, 2006; revised on June 19, 2006; accepted on June 23, 2006
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Abstract |
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The carbohydrates present on the surface of differentiated human embryonic stem cells (hESCs) are not yet well established. Here, we have employed a panel of lectins and several anti-carbohydrate antibodies to determine the carbohydrates that are present at day 12 of hESC differentiation as embryoid bodies (EBs). On the basis of staining with fluorescein-labeled lectins, we have determined the presence of both terminal and internally linked
-D-mannopyranosyl groups, poly-N-acetyllactosaminyl chains, both 2,3- and 2,6-linked N-acetylneuraminic acid (Neu5Ac), 1,6-linked L-fucosyl, and ß-D-galactosyl groups, and more specifically, the T, Tn, and sialyl-Tn antigens. However, no 1,2-linked L-fucosyl, terminal nonreducing -D-galactosyl, N-acetyl-ß-D-glucosaminyl, nor N-acetyl--D-galactosaminyl groups were found by this approach. We also established the presence of Neu5Ac2,3/2,6-Galß1,4 GlcNAc-terminated chains on the surfaces of 12-day-old EBs, as indicated by the great enhancement of staining by Erythrina cristagalli agglutinin (ECA) after treatment with neuraminidase. In each case, inhibition of binding by a haptenic sugar or treatment with neuraminidase was used to eliminate the possibility of nonspecific binding of the lectins. A comparison with undifferentiated cell staining revealed an increase in 2,3-linked Neu5Ac as well as a change to exclusively 1,6-linked L-fucose upon differentiation. Key words: carbohydrates / differentiation / embryoid body / human embryonic stem cells / lectin
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Introduction |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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Cell surface carbohydrates are importantly involved in cell–cell interaction and communication, cell adhesion, and many developmental processes. The question of the function of cell surface glycans during the early development of organisms is an important area for investigation as these glycans may play a role in differentiation (Haltiwanger and Lowe, 2004
). Several studies have identified antigenic glycoproteins and glycolipids in the earliest development of the embryo. Two of these antigens, SSEA-3 and SSEA-4, have been identified as appearing during the four- to eight-cell stage of mouse embryos and human teratocarcinoma cell lines (Kannagi et al., 1982; Shevinsky et al., 1982). Two obvious tools with which to identify and map cell surface carbohydrates are lectins and antibodies. These probes, used with appropriate controls (blocking sugar ligands and glycosidases), can provide important information on the presence and changes during development of sugar residues on the surface of stem cells and differentiating embryoid bodies (EBs). A recent study on undifferentiated human embryonic stem cells (hESCs) used lectins to establish the presence of sugar residues on the surface of BG01 and BG02 undifferentiated hESCs (Venable et al., 2005). The present work employs many more lectins and carbohydrate antibodies to identify the presence of carbohydrates on the surface of differentiating EBs. Using the same methodology, we compared our results on differentiated EBs with those on undifferentiated cells. |
Results |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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We employed 34 fluorescein-labeled lectins1 (Goldstein and Poretz, 1986
; Goldstein et al., 1997; Sharon and Lis, 2003) and several monoclonal anti-carbohydrate antibodies to assess carbohydrates expressed by EBs. In all cases, we established specificity of lectin and antibody binding with the use of specific carbohydrate inhibitors of binding and/or specific glycosidases, for example, neuraminidase to abolish binding by the elderberry bark lectin (Sambucus nigra agglutinin, SNA). Table I summarizes the lectins and antibodies used in this study along with data for the conditions employed in their use such as concentration used for staining and sugar haptens and glycosidases used to establish specificity of lectin binding.
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High amounts of -mannopyranosyl2 groups are present as indicated by strong staining with concanavalin A (Con A) and the banana lectin (BanLec, Musa acuminata) (Figure 1) and moderate staining by the snowdrop bulb lectin (Galanthus nivalis agglutinin, GNA). All recognize terminal nonreducing -mannosyl groups, whereas Con A and BanLec also recognize glucose and the internal branched Man3 core trisaccharide (Man -1,6[Man 1–3] Man) of N-linked glycans. The BanLec also can interact with internal 1,3-linked Man groups. In all cases, lectin binding was abolished in the presence of the specific inhibitor, methyl -mannopyranoside.
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The presence of poly-N-acetyllactosaminyl chains (LacNAc) is clearly demonstrated by the intense staining by fluorescein-labeled Erythrina cristagalli agglutinin (ECA), wheat germ agglutinin (WGA, Triticum vulgaris), and succinylated WGA (WGA-succ), tomato lectin (Lycopersicon esculentum agglutinin, LEA) (Figure 2), potato lectin (Solanum tuberosum lectin, STL), and jimson weed lectin (Datura stramonium agglutinin, DSA), all of which strongly recognize this carbohydrate epitope. Inhibition of staining using LacNAc or N,N',N''-triacetylchitotriose established specificity of binding by these lectins. It should be noted that although several of these lectins (e.g., WGA, LEA, and STL) are most often described as GlcNAc-dextrin-binding lectins, they also react with LacNAc and its homologs (Kawashima et al., 1990). Consistent with the presence of terminal N-acetyllactosaminyl residues is the staining by Ricinus communis agglutinin (RCA) and Ischnoderma resinosum lectin (IRL) (Figure 3), both terminal ß-galactosyl-recognizing lectins. Staining with ECA was strongly enhanced following treatment with neuraminidase, indicating the presence of Neu5Ac 2,3/2,6-Gal ß1,4GlcNAc-terminated chains (Figure 4).
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Twelve-day differentiated hESCs exhibit both the Tn (GalNAc -O-Ser/Thr) (Figure 5) and sialyl-Tn (Neu5Ac -2,6-GalNAc -O-Ser/Thr) (Figure 6) antigens. This is indicated by positive staining with soybean agglutinin (SBA, Glycine max) and anti-Tn antibody and by SNA and anti-sialyl-Tn antibody, respectively. Prior treatment of hESCs with neuraminidase abolished the reaction with anti-sialyl-Tn antibody. The presence of the T-disaccharide (Gal ß1,3 GalNAc) was shown by staining with the peanut lectin (PNA, Arachis hypogaea), amaranthin (Amaranthus caudatus agglutinin, ACA), and the Agaricus bisporus agglutinin (ABA) mushroom lectin.
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The presence of sialic acid is clearly indicated by staining by the following lectins: slug (Limax flavus agglutinin, LFA), horseshoe crab (Limulus polyphemus agglutinin, LPA), Maackia amurensis agglutinin (MAA) (specific for 2,3-linked Neu5Ac), SNA (specific for Neu5Ac2,6Gal/GalNAc) (Figure 7), and Polyporus squamosus lectin (PSL), which recognizes sialic acid linked 2,6 to N-acetyllactosamine, a trisaccharide epitope that occurs only on N-linked chains of glycoproteins. The presence of trisialyl N-glycans was indicated by strong staining by the Psathyrella velutina mushroom lectin (PVL) (Ueda et al., 2002). Neu5Ac abolished labeling by the slug and horseshoe crab lectins; 2,3-sialyllactosamine blocked MAA; and the treatment with neuraminidase negated staining by SNA, LFA, PVL, and PSL. Staining by WGA, which also recognizes sialic acid bound to LacNAc, is diminished by treating cells with neuraminidase.
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Of many fucosyl-binding lectins assayed for binding to hESCs, only the orange peel fungal lectin (Aleuria aurantia lectin, AAL) stained the cells (Figure 8). This lectin is known to recognize 1,6-linked fucosyl (see footnote 2) groups as occur on the N,N'-diacetylchitobiosyl residues in N-linked glycoproteins (Fukumori et al., 1990). The lotus lectin (Lotus tetragonolobus lectin, LTL) and UEA1 (Ulex europaeus), which bind principally to 1,2-linked L-fucosyl groups, did not stain the cells nor did the eel serum fucosyl-binding lectin (Anguilla anguilla agglutinin, AAA).
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Lectins reacting with nonreducing terminal -GalNAc on oligosaccharides, including the lima bean lectin (LBL) (Phaseolus lunatus agglutinin, PLA), Dolichos biflorus agglutinin (DBA), and the snail, Helix pomatia agglutinin (HPA) failed to bind. The soybean lectin, also an -GalNAc-binding lectin, gave a positive result; it alone, of all the GalNAc-binding lectins tested, most readily recognizes a single GalNAc residue O-linked to Ser/Thr. Terminal nonreducing ß-GlcNAc groups also appear to be absent from 12-day-old EBs, from the observation that the Griffonia simplicifolia II (GS II) lectin did not bind. We did not detect significant amounts of terminal -galactosyl groups by the Griffonia simplicifolia I B4 (GS I-B4) isolectin; however, slight binding by Marasmius oreades agglutinin (MOA), which is highly specific for the Gal 1,3Gal ß1,4GlcNAc/Glc epitope, was apparent. LacNAc reacts very weakly with MOA, and, given the presence of extensive LacNAc termini as shown by the binding of ECA, LEA, WGA, STL, and DSA, this is a possible explanation for its weak binding.
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Discussion |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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This study provides an in-depth description of the carbohydrates on the surface of 12-day differentiated EBs as revealed by the use of carbohydrate-recognizing proteins: lectins and anti-carbohydrate antibodies. Our results indicate the presence of (1) extensive
-mannosyl-terminated chains as shown by moderate staining by GNA and intense staining by Con A and the BanLec, the last two of which also recognize the branched Man3 trisaccharide proximal to Asp-linkage in N-linked glycoproteins; (2) poly N-acetyllactosaminyl residues indicated by staining with ECA, WGA, and the tomato, potato, and jimson weed lectins. Staining by RCA and IRL confirms the presence of terminal ß-galactosyl groups; (3) sialylated glycoconjugates as revealed by staining by LFA, LPA, SNA, PVL, and PSL, all of which recognize 2,6-linked Neu5Ac, and by MAA, which binds 2,3-linked sialic acid; (4) 1,6-linked fucose as shown by the binding of the orange peel fungal lectin, AAL. This fucosyl group is probably on the N,N'-diacetylchitobiosyl residue linked to an Asn residue of glycoprotein; (5) Tn and sialyl-Tn antigens as revealed by staining by SBA and antibodies against these carbohydrate antigens; and (6) The T-(Thomsen-Friedenreich) antigen (Gal 1,3GalNAc-O-Ser/Thr) as revealed by staining by PNA, ACA, and ABA. On the contrary, lack of lectin staining suggests the absence of the following carbohydrate residues: terminal -galactosyl, N-acetylglucosaminyl, and N-acetylgalactosaminyl groups as well as the probable lack of 1,2-linked fucosyl groups. Many of the lectins stain only the cell surface and/or cytoplasm of the ES cells. This is demonstrated by BanLec (Figure 1), LEA (Figure 2), and IRL (Figure 3). Other lectins appear to stain all parts of the cells—nucleus, cytoplasm, and cell surface—and entire EBs, for example, SNA (Figure 7) and AAL (Figure 8). Still other lectins stain entire cells, but only those located at the edges or in sporadic locations of the EBs. ECA (Figure 4) is an example of a lectin that normally stains only edges and sporadic locations within an EB. Following neuraminidase treatment of the cells, this lectin appears to stain all cell surfaces and cytoplasm but not nuclei. No correlation is evident between specificity and location of stain because different lectins having the same general specificity display different binding locations.
To compare 12-day differentiated EBs with undifferentiated hESCs, we used similar fluorescence microscopy techniques on fixed hESCs rather than relying solely on previous data from cell-sorting techniques (Venable et al., 2005) (Table II). It is interesting to note, however, that the data achieved by the two methods were very similar. Using our own undifferentiated cell data, we were able to draw the following conclusions: MAA bound 89% of 12-day EBs, whereas only 19% of the undifferentiated cells were labeled by this lectin. This binding by MAA is also consistent with the presence in human EBs of the stage-specific embryonic antigens SSEA-3 and SSEA-4, the globogangliosides occurring in human teratocarcinoma cells (Kannagi et al., 1982). AAL bound to 100% of both undifferentiated and 12-day differentiated cells, indicating the presence of 1,2- and/or 1,6-linked fucose in both cell types. We employed UEA1 to determine which type of fucose was present in each type of cells. UEA1 stained 15% of the undifferentiated cells but none of the 12-day differentiated cells, indicating loss of virtually all 1,2-linked fucose during the early stage of differentiation. To our knowledge, this is the first time that such a shift has been identified. We believe that these changes in undifferentiated hESCs during the 12-day differentiation as EBs may be of importance.
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We also identified for the first time the presence of the Tn and sialyl-Tn antigens in 12-day-old EBs. As positive controls, we observed positive staining of the sialyl-Tn antibody to human cervix, testis, and duodenum tissues and for the Tn antibody to the cervix and duodenum tissues (Yonezawa et al., 1992). The presence of the T-disaccharide was also shown by the binding of ABA and PNA, as well as the very strong staining by ACA, all of which recognize this carbohydrate epitope (Springer, 1984).
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Materials and methods |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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Materials
The lectins BanLec, Con A, DSA, GNA, IRL, LBL, LFA, MAA, MOA, PSL, and SNA were prepared from their natural sources and labeled with fluorescein isothiocyanate (FITC) (Sigma, St. Louis, MO) in this laboratory. The following lectins were purchased from EY laboratories (San Mateo, CA) as FITC-conjugated: AAA, ABA, GS II, HPA, LPA, PHA-E (Phaseolus vulgaris erythroagglutinin), PHA-L, (Phaseolus vulgaris leucoagglutinin), PNA, and UEA1. The lectin AAL was purchased from Vector laboratories (Burlingame, CA) and was FITC-labeled following the procedure described below. The following lectins were purchased from Vector laboratories as FITC-labeled: ACA, DBA, ECA, EEA (Euonymus europaeus agglutinin), GS I-B4, LEA, LTL, RCA1, SBA (Glycine max), STL, WGA, and WGA-succ. One lectin, PVL, was purchased from WAKO Chemicals (Richmond, VA) and FITC-labeled in this laboratory. Before use, all lectins were checked by spectrophotometry for FITC-labeling, and, if possible, an agglutination assay was done to assure specific binding ability. The only lectin that did not demonstrate agglutinating activity, LBL, is thought to have been split into its monomers during FITC labeling. It is presumed to retain its specific binding ability, although it no longer agglutinates red blood cells (RBCs). The following haptenic sugars were purchased from Sigma: Neu5Ac (used to block LFA and LPA),
2,3 sialyllactose (used to block MAA), lactose (used to block PSL, IRL, PNA, and RCA), fucose (used to block AAL), and the benzyl T-antigen (used to block ABA and ACA). Human milk sialyllactose (80% 2,6-linked and 20% 2,3-linked), used to block SNA, was also purchased at Sigma. The N-acetylgalactopyrannoside used to block SBA and the N-acetylglucopyrannoside used to block PVL were purchased from Pfanstiehl Laboratories (Waukegan, IL). The Gal 1,3 Gal used to block MOA was purchased from V-labs, Inc., (Covington, LA). The remaining sugars used in this study, methyl -mannoside (used to block BanLec, Con A, and GNA), N,N',N''-triacetylchitotriose (used to block DSA, LEA, STL, and WGA-succ), and LacNAc (used to block WGA and ECA), were all available in this laboratory from previous studies. The anti-Tn antigen, sialylated mouse mAb antibody (clone B35.2—reacts only with NeuAc2,6GalNAc-Ser/Thr), was purchased from Cal Bio Chem (La Jolla, CA). The monoclonal mouse antibody to the Tn antigen (clone B1.1) was purchased from Biomeda (Foster City, CA), and the secondary antibody, Cy3-conjugated donkey anti-mouse, was purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). Clostridium perfringens neuraminidase was purchased from Sigma as a lyophilized powder. Coverslips used in undifferentiated cell culture were 18 mm in diameter and between 0.13 and 0.17 mm thick. The coverslips were purchased from Fisher (Marshalltown, IA). The Triton X-100 was purchased from Research Products International (Elk Grove Village, IL).
Fluorescein labeling of lectins
One to four milligrams of the desired lectin was placed into a dark vial and dissolved in 1 mL of phosphate-buffered saline (PBS, 2 mM sodium phosphate monobasic, 8 mM sodium phosphate dibasic, 0.15 M sodium chloride, 0.1 mM CaCl2, and 6 mM sodium azide) containing 100 mM sodium carbonate at pH 9.5. Lectin-specific sugars were added to the solution to protect the active sites, mixed briefly, and allowed to stand for at least 10 min to ensure complete binding to the active site. To this solution was added 
1 mg FITC. The vial was rotated in the dark for 1–2 h at room temperature depending on the lectin being labeled, after which the solution was transferred to a dialysis bag. Dialysis into PBS proceeded in the dark until all unbound FITC and carbohydrate were removed. The solution was stored in an amber vial at 4°C until use.
Undifferentiated hESC culture
BG01 hESCs were cultured in vitro on a mouse embryonic fibroblast (MEF) feeder layer in hESC medium (78.7% Dulbecco’s modified Eagle’s medium [DMEM; Invitrogen, Carlsbad, CA], 19.7% knockout serum replacement [KOSR; Invitrogen], 1.0% nonessential amino acids [NEAA; Invitrogen], 0.2% basic fibroblast growth factor [bFGF; Invitrogen], 0.1 mM ß-mercaptoethanol [Sigma], and 2 mM L-glutamine [Invitrogen]). Two confluent 60-mm hESC dishes were used for each 12-well plate of undifferentiated cells. A round 18-mm diameter coverslip 0.13–0.17 mm thick (Fisher) was placed into each well, and undifferentiated hESCs were plated on top of these.
Lectin staining of undifferentiated hESCs
Between 2 and 4 days after plating, the hESCs were fixed in a solution of 2% paraformaldehyde and 0.1% Triton X-100 in PBS (2 mM sodium phosphate monobasic, 8 mM sodium phosphate dibasic, 0.15 M sodium chloride, 0.1 mM CaCl2, and 6 mM sodium azide). The fixed cells were then rinsed three times in PBS-containing 0.1% Triton X-100 for 5 min each. The cells were placed in a dark chamber, and a solution of 1% bovine serum albumin (BSA, Sigma), 0.1% Triton X-100 in PBS, was applied for 20 min, followed by treatment for 20 min with a solution in PBS-containing lectin at the specified concentration and 0.1% Triton X-100. The cells were rinsed three times for 5 min each with a solution of 0.1% Triton X-100 in PBS and stained with a solution of 1 x Hoechst 23187 (Sigma) and 0.1% Triton X-100 in PBS for 5 min. Following rinsing the cells once with PBS, the coverslips were removed from the wells, rinsed once by dipping into PBS, and applied directly to a slide containing 7–10 µL of Prolong Gold (Molecular Probes, Eugene, OR). After standing in the dark chamber overnight, the slides were stored in the dark at –20°C until they were viewed on a fluorescence microscope.
EB formation from hESCs
BG01 hESCs were cultured in vitro on an MEF feeder layer before differentiation. Two confluent 60-mm hESC dishes were used for each dish of EBs prepared. The cells that had already differentiated were removed from the dishes, leaving only pluripotent hESCs in hESC medium. To generate EBs, undifferentiated hESCs were removed from the MEF feeder, then suspended in 10 mL hESC medium, and centrifuged at 78 x g for 5 min. The cells were resuspended in EB medium made of 78.8% DMEM, 19.7% KOSR, 1.0% NEAA, 0.1 mM ß-mercaptoethanol, and 2 mM L-glutamine and placed in a dish to form EBs. The EB medium was replaced every other day for 12 days.
Cryostat embedding and sectioning
Cryomolds (Electron Microscopy Sciences, Hatfield, PA) were filled with optimum cooling temperature (OCT) medium (TissueTek; Electron Microscopy Sciences). The 12-day EBs were placed in a 15-mL conical vial, centrifuged briefly, and the supernatant was removed. The EBs were then rinsed with 5-mL room temperature 1 x PBS (Invitrogen) and placed in a 1.5-mL tube. The tube was centrifuged briefly and the PBS removed. The pelleted EBs were removed using a glass pipet, transferred to the mold containing the OCT medium, and then frozen in a 2-methylbutane dry ice slurry. Samples were stored at –80°C until sectioning. Six-micrometer sections were cut using a Microm cryostat and collected on positively charged slides (Fisher), which were stored at –80°C until staining.
Lectin staining of differentiated cryostat sections
After thawing, samples were outlined using a pap pen (Electron Microscopy Sciences). They were rinsed once with PBS for 5 min, blocked in a solution of 1% BSA in PBS for 20 min followed by lectin solution in PBS for 20 min. The samples were rinsed three times in PBS for 5 min each and fixed in a 2% solution of paraformaldehyde in PBS for 15 min. After fixation, the samples were rinsed three times in PBS for 5 min each. A 1 x Hoechst 23187 solution (Sigma) in PBS was applied as a nuclear stain for 5 min. The slides were further rinsed one time with PBS for 5 min. Slides were coverslipped with 5–10 µL Prolong Gold (Molecular Probes) and stored at –20°C until they were viewed on an Olympus BX51 fluorescent microscope. Unless otherwise specified, all procedures were completed in low light at room temperature in a moist chamber. Control slides were treated with PBS or lectin solution incubated with the blocking carbohydrate for at least 20 min in place of the lectin solution. In the case of LPA, HEPES buffer (0.15 M sodium chloride, 0.02 M HEPES, 0.01 M calcium chloride, and 0.04% sodium azide) was used to prepare all reagents in place of PBS because LPA requires a high concentration of calcium to bind.
Antibody staining
After thawing, frozen sections were outlined with a pap pen. They were rinsed once with PBS for 5 min before applying a blocking buffer of 1% donkey serum (Jackson ImmunoResearch Laboratories) and 0.5% Triton X-100 in PBS for 1 h. After blocking, the primary antibodies were diluted in blocking buffer and applied to the slides overnight at 4°C. The slides were then rinsed three times in PBS for 5 min. The secondary antibodies were diluted in the blocking buffer and applied to the samples for 1 h at room temperature, then rinsed three times with PBS. As necessary, the lectin procedure was followed from this point on, beginning with BSA treatment of the slide wells. For controls, blocking buffer was used in place of primary antibodies. Unless otherwise specified, all procedures were completed in diminished light at room temperature in a moist chamber.
Neuraminidase treatment
Lyophilized neuraminidase was reconstituted to a concentration of 5-units/mL in 0.1 M sodium acetate buffer at pH 5.0 containing 0.15 M sodium chloride. This solution was diluted 5-fold in the same buffer and applied to the well. The slide was placed in a moist chamber and incubated at 37°C for 1 h, after which it was brought to room temperature and rinsed three times in PBS for 5 min each. The slide was then stained by antibody or lectin following the procedure described above.
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Conflict of interest statement |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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None declared.
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Acknowledgments |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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We thank Crystal Pacut for culturing the hESCs and providing the slides used in this project. We also acknowledge support from the NIH (grants GM29470 and p20GM069985).
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Footnotes |
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1 The authors have not provided references for the carbohydrate-binding specificity of each lectin employed in this article. Rather they have cited several monographs, which provide the information on the carbohydrate-binding properties of the lectins employed in this study. When appropriate, citations that provide more recent or relative information are noted.
2 All carbohydrates reported in this article are of the D-configuration except for fucose, which is of the L-configuration.
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Abbreviations |
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AAA, Anguilla anguilla agglutinin, eel serum; AAL, Aleuria aurantia lectin, orange peel fungus; ABA, Agaricus bisporus agglutinin; ACA or amaranthin, Amaranthus caudatus agglutinin; BanLec, banana lectin, Musa acuminata; BSA, bovine serum albumin; Con A, concanavalin A; DBA, Dolichos biflorus agglutinin; DSA, Datura stramonium agglutinin, jimson weed; EB, embryoid body; ECA, Erythrina cristagalli agglutinin; EEA, Euonymus europaeus agglutinin; FITC, fluorescein isothiocyanate; GNA, Galanthus nivalis agglutinin, snowdrop bulb lectin; GS I-B4, Griffonia simplicifolia I B4; GS II, Griffonia simplicifolia II; hESC, human embryonic stem cell; HPA, Helix pomatia agglutinin, edible snail; IRL, Ischnoderma resinosum lectin; LacNAc, N-acetyllactosamine; LBL, lima bean lectin; LEA, Lycopersicon esculentum agglutinin, tomato lectin; LFA, Limax flavus agglutinin, slug; LPA, Limulus polyphemus agglutinin, horseshoe crab lectin; LTL, Lotus tetragonolobus lectin; MAA, Maackia amurensis agglutinin; MEF, mouse embryonic fibroblast; MOA, Marasmius oreades agglutinin; Neu5Ac, N-acetylneuraminic acid; PBS, phosphate buffered saline; PHA-E, Phaseolus vulgaris erythroagglutinin; PHA-L, Phaseolus vulgaris leucoagglutinin; PLA, Phaseolus lunatus agglutinin; PNA, peanut agglutinin, Arachis hypogaea; PSL, Polyporus squamosus lectin; PVL, Psathyrella velutina lectin; RCA, Ricinus communis agglutinin; SBA, Glycine max, soybean agglutinin; SNA, Sambucus nigra agglutinin, elderberry bark lectin; SSEA, stage-specific embryonic antigen; STL, Solanum tuberosum lectin, potato lectin; UEA1, Ulex europaeus agglutinin; WGA, wheat germ agglutinin, Triticum vulgaris; WGA-succ, succinylated WGA.
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References |
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TopAbstractIntroductionResultsDiscussionMaterials and methodsConflict of interest statementAcknowledgmentsReferences |
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Fukumori, F., Takeuchi, N., Hagiwara, T., Ohbayashi, H., Endo, T., Kochibe, N., Nagata, Y., and Kobata, A. (1990) Primary structure of a fucose-specific lectin obtained from a mushroom, Aleuria aurantia. J. Biochem., 107, 190–196.
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Venable, A., Mitalipova, M., Lyons, I., Jones, K., Shin, S., Pierce, M., and Stice, S. (2005) Lectin binding profiles of SSEA-4 enriched, pluripotent human embryonic stem cell surfaces. BMC Dev. Biol., 5, 15.[CrossRef][Medline]
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S.-C. Tao, Y. Li, J. Zhou, J. Qian, R. L Schnaar, Y. Zhang, I. J Goldstein, H. Zhu, and J. P Schneck Lectin microarrays identify cell-specific and functionally significant cell surface glycan markers Glycobiology, October 1, 2008; 18(10): 761 - 769. [Abstract] [Full Text] [PDF]
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