Structural basis for altered soybean agglutinin lectin binding between a murine metastatic lymphoma and an adhesive low malignant variant

Lang, Elke; Köhl, Ulrike; Schirrmacher, Volker; Brossmer, Reinhard; Altevogt, Peter

doi:10.1016/0014-4827(87)90348-x

Cited by 22 publications

(25 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The staining of cells with hybridoma supernatant-and PE-conjugated goat antibodies to rat immunoglobulins (SERVA, Heidelberg, FRG) has been described in detail elsewhere (41). Stained cells were analyzed with a FACScan | fluorescenceactivated cell analyzer (Becton Dickinson, Heidelberg, FRG).…”

Section: Methodsmentioning

confidence: 99%

“…Lactoperoxidase-catalyzed iodination of surface glycoproteins on intact cells, cell lysis in the presence of NP-40, immunoprecipitation using protein G-Sepharose or protein A/MAR, treatment of precipitates with endoglycosidase F/Nglycosidase F (Boehringer Mannheim, Mannheim, FRG) has been Homophilic Binding of a4 Integrins described elsewhere (41). SDS-PAGE was performed on 10% slab gels.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The alpha 4 integrin chain is a ligand for alpha 4 beta 7 and alpha 4 beta 1.

Altevogt

Hubbe

Ruppert

et al. 1995

The Journal of Experimental Medicine

View full text Add to dashboard Cite

SummaryThe heterodimeric ol4 integrins c~4~7 lymphocyte Peyer's patch adhesion molecule ([LPAM]-I) and ol431 (very late antigen-4) are cell surface adhesion molecules involved in lymphocyte trafficking and lymphocyte-cell and matrix interactions. Known cellular ligands include vascular cell adhesion molecule (VCAM)-I, which binds to ~431 and ol437, and the mucosal addressin cell adhesion molecule (MAdCAM)-I, which binds to ol437. Here we show that the o~4 chain of these integrins can itself serve as a ligand. The c~4 chain, immunoaffinity purified and immobilized on glass slides, binds thymocytes and T lymphocytes. Binding exhibits divalent cation requirements and temperature sensitivity which are characteristic of integrin-mediated interactions, and is specifically inhibited by anti-cx4 integrin antibodies, which exert their effect at the cell surface. Cells expressing exclusively c~4/~7 (TK-1) or c~431 (L1-2) both bound avidly, whereas ol4-negative cells did not. A soluble 34-kD ol4 chain fragment retained binding activity, and it inhibited lymphocyte adhesion to ~4 ligands. It has been shown that c~4 integrin binding to fibronectin involves an leucine-aspartic acid-valine (LDV) motif in the HeplI/IIICS region of fibronectin (CS-1 peptide), and homologous sequences are important in binding to VCAM-1 and MAdCAM-1. Three conserved LDV motifs occur in the extracellular sequence of c~4. A synthetic LDV-containing c~4-derived oligopeptide supports c~4-integrin-dependent lymphocyte adhesion and blocks binding to the 34-kD ol4 chain fragment. Our results suggest that r and r integrins may be able to bind to the ol4 subunit on adjacent cells, providing a novel mechanism for ol4 integrin-mediated and activation-regulated lymphocyte interactions during immune responses.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The alpha 4 integrin chain is a ligand for alpha 4 beta 7 and alpha 4 beta 1.

Altevogt

Hubbe

Ruppert

et al. 1995

The Journal of Experimental Medicine

View full text Add to dashboard Cite

show abstract

“…The detailed procedure for indirect immunofluorescence with mAb has been described in previous publications [27]. Fluorescein isothiocyanate (F1TC)-conjugated F(ab')z fragments of affinity-purified goat anti-rat Ig (Dianova, Hamburg, FRG) were used as second antibody.…”

Section: Indirect Immunofluorescence Staining Of Cellsmentioning

confidence: 99%

Antibody 12‐15 cross‐reacts with mouse Feγ receptors and CD2: study of thymus expression, genetic polymorphism and biosynthesis of the CD2 protein

et al. 1989

Self Cite

View full text Add to dashboard Cite

Previously we described a monoclonal antibody (mAb 12-15) that reacted with murine Fc receptor proteins (beta 1, beta 2 and alpha) and an undefined molecule of 37 kDa (beta 3) on certain types of cells. Here we present serological and biochemical evidence that the beta 3 chain is expressed on mouse thymocytes and that it is identical to the CD2 antigen. By immunofluorescence staining and cytofluorographic analysis greater than 95% of thymocytes were positive. Brightly staining cells coincided with cortisone-resistant thymocytes suggesting that mature thymocytes expressed higher levels of the antigen. Biosynthetic labeling of DBA/2 thymocytes with [35S]methionine showed that the size of the CD2 precursor molecule was 43 kDa which was processed to approximately 55-65 kDa in the mature molecule. mAb 12-15 was also reactive with the tunicamycin-treated form of the CD2 antigen suggesting that the cross-reactive epitope was of protein nature. Comparison of different mouse strains indicated that two molecular forms of CD2 exist. On BALB/c thymocytes, the relative mass of the native molecule was approximately 60-70 kDa (CD2.1) and slightly larger than in DBA/2 (CD2.2). Following endoglycosidase F treatment both proteins still showed a slight difference in electrophoretic mobility. Several inbred mouse strains were analyzed for expression of CD2 forms. When mAb 12-15 was used in cytotoxic T lymphocyte inhibition experiments using specific CTL and tumor target cells it was found that the antibody could specifically inhibit CTL-mediated lysis presumably by interfering with CD2 function.

show abstract

“…Cytofluorography-The cell-surface staining of cells with saturating amounts of mAbs, either hybridoma supernatants or purified antibodies, and phycoerythrin-conjugated goat antibodies to rat Ig (SERVA, Heidelberg, FRG) or hamster Ig (Dianova, Hamburg, FRG) has been described elsewhere (39). For the detection of cytoplasmic staining cells were first incubated on ice with 0.5% formaldehyde in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…Biochemical Analysis-Lactoperoxidase-catalyzed iodination of intact cells was carried out as described (39). Following the labeling reaction, the cells were washed three times in PBS and lysed at 4°C for 15 min in Tris-buffered saline containing either 0.3% CHAPS (Sigma), 1% IGEPAL CA-630 (Sigma) or 1% IGEPAL CA-630 in the presence of 2 mM Ca 2ϩ and Mg 2ϩ ions.…”

Section: Methodsmentioning

confidence: 99%

Asp-698 and Asp-811 of the Integrin α4-Subunit Are Critical for the Formation of a Functional Heterodimer

Zeller¹,

Lohr²,

Sammar³

et al. 1998

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The amino acid motif LDV is the principal binding site for ␣ 4 integrins in fibronectin, and homologous motifs are recognized in vascular cell adhesion molecule-1 and MAdCAM-1. Three conserved LDV motifs (LDV-1 to 3) occur in the ectodomain of the human and mouse ␣ 4 -subunit, the functions of which are unknown. We demonstrate here that ␣ 4 -transfected fibroblasts with mutation in LDV-1 (D489N) behaved like ␣ 4 -wild type but that LDV-2 (D698N) and LDV-3 (D811N) mutants were impaired in binding and spreading on ␣ 4 -specific substrates. On the RGD-containing fibronectin fragment FN-120 there was an inverse behavior; now the ␣ 4 -wild type and the LDV-1 mutant could not adhere whereas the two other mutants could. The

show abstract

Structural basis for altered soybean agglutinin lectin binding between a murine metastatic lymphoma and an adhesive low malignant variant

Cited by 22 publications

References 27 publications

The alpha 4 integrin chain is a ligand for alpha 4 beta 7 and alpha 4 beta 1.

The alpha 4 integrin chain is a ligand for alpha 4 beta 7 and alpha 4 beta 1.

Antibody 12‐15 cross‐reacts with mouse Feγ receptors and CD2: study of thymus expression, genetic polymorphism and biosynthesis of the CD2 protein

Asp-698 and Asp-811 of the Integrin α4-Subunit Are Critical for the Formation of a Functional Heterodimer

Contact Info

Product

Resources

About