The epidermal growth factor–like domains of the human EMR2 receptor mediate cell attachment through chondroitin sulfate glycosaminoglycans

Stacey, Martin; Chang, Gin-Wen; Davies, John Q.; Kwakkenbos, Mark J.; Sanderson, Ralph D.; Hamann, Jörg; Gordon, Siamon; Lin, Hsi‐Hsien

doi:10.1182/blood-2002-11-3540

Cited by 195 publications

(225 citation statements)

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“…17,39 Other than CD97, EMR2 does not recognize CD55. 12,39 Three different amino acids within the first 2 EGF domains account for this difference in ligand binding. 11 The first different amino acid is Asn 33 in CD97, one of the N-glycosylation sites within the first EGF domain of CD97.…”

Section: Discussionmentioning

confidence: 99%

“…9 -11 Chondroitin sulfate glycosaminoglycan, present in cell membranes and the extracellular matrix, is a new ligand to the longest CD97 isoform (EGF 1,2,3,4,5). 12 The ability of CD97 EGF domains to interact with cellular and/or extracellular matrix ligands suggests that the molecule is involved in cell-cell or cell-matrix interactions. Various lines of evidence have shown an important role for CD97 in tumor dedifferentiation, migration, invasiveness and metastasis.…”

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confidence: 99%

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N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

Wobus

Vogel

Schmücking

et al. 2004

Intl Journal of Cancer

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CD97 is an EGF-TM7 receptor found on various carcinomas where expression levels correlate with dedifferentiation and tumor stage, smooth muscle cells and leukocytes. CD97 acts as an adhesion molecule by binding to its cellular ligand, CD55. In this study, we demonstrate that 2 immunodominant CD97 epitopes are not equally present in the various cell types. Differences were apparent in gastrointestinal tumors and smooth muscle cells where monoclonal antibodies (mAbs) to the first epidermal growth factor (EGF) domain (CD97 EGF ) showed a more restricted staining pattern than mAbs to the stalk region (CD97 stalk ). This discrepancy was not detectable in cultured gastrointestinal tumor cell lines. In fact, the selection of the CD97 mAb influences the result of clinical studies. Thus, we clarified the reason(s) for these differences in CD97 mAb staining on various cell types. We provide evidence that epitope accessibility for CD97 EGF Key words: CD97; colorectal cancer; smooth muscle cell; epitope accessibility; glycosylation CD97 is a member of a subfamily of 7-span transmembrane (TM7) receptors referred to as EGF-TM7 proteins. 1,2 In normal tissues, CD97 is abundantly expressed in smooth muscle cells, macrophages, granulocytes, dendritic cells and in some T and B cells. 3 Carcinomas of different origin showed higher CD97 expression compared to the corresponding normal cell types. 4 -7 CD97 consists of an extended extracellular region with several N-terminal epidermal growth factor (EGF) domains coupled to the TM7 domain by a stalk. 1,8 Various CD97 isoforms exist possessing 3 (EGF 1,2,5), 4 (EGF 1,2,3,5), or 5 (EGF 1,2,3,4,5) EGF domains. CD97 binds to membrane receptor CD55. 9 This interaction is mediated by the first 2 CD97 EGF domains. 9 -11 Chondroitin sulfate glycosaminoglycan, present in cell membranes and the extracellular matrix, is a new ligand to the longest CD97 isoform (EGF 1,2,3,4,5). 12 The ability of CD97 EGF domains to interact with cellular and/or extracellular matrix ligands suggests that the molecule is involved in cell-cell or cell-matrix interactions. Various lines of evidence have shown an important role for CD97 in tumor dedifferentiation, migration, invasiveness and metastasis. CD97 levels correlate with the in vitro migration and invasion capacity of colorectal tumor cell lines and CD97-transfected cells. 6 Colorectal carcinomas with more strongly CD97-stained, scattered tumor cells at the invasion front showed a poorer clinical stage as well as increased lymph vessel invasion compared to cases with uniform CD97 staining. 6 In thyroid cancers, CD97 expression parallels the aggressiveness and lymph node involvement of these tumors. 4,5 Our previous data revealed that CD97 has prognostic and therapeutic potential in cancer. However, analyzing CD97 expression brings up the phenomenon that different staining patterns, depending on which CD97 monoclonal antibody (mAb) has been used for immunohistology, can be readily observed in pancreatic tissues. 7,13 Based on this observation, one aim of ...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

Wobus

Vogel

Schmücking

et al. 2004

Intl Journal of Cancer

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“…Together with CD97 and EMR2 through EMR4, it belongs to the EGF-TM7 family, a subfamily within the adhesion class of TM7 receptors [10,11]. All these receptors have variable numbers of EGF-like domains, which can mediate ligand binding [12,13]. Although a cellular ligand might be anticipated, molecules interacting with F4/80 have not yet been identified.…”

Section: Introductionmentioning

confidence: 99%

EMR1, the human homolog of F4/80, is an eosinophil‐specific receptor

et al. 2007

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The EGF-TM7 F4/80 is a defining marker of murine macrophage populations. Applying flow cytometric analysis using the newly generated mAb A10, and quantitative real-time PCR, we here report the surprising observation that the human ortholog of F4/80, EGFlike module containing mucin-like hormone receptor (EMR)1, is absent on mononuclear phagocytic cells including monocytes, macrophages, and myeloid dendritic cells. Unexpectedly, we found that EMR1 expression is restricted to eosinophilic granulocytes, where expression is overlapping with the eotaxin receptor CCR3 and the immunoglobulin-like lectin Siglec-8. Absence on other leukocytes, including basophils, implies that EMR1 is a highly specific marker for eosinophils in humans.

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“…[7][8][9] The coding sequences for cbEGF [7][8][9] (amino acids 300-429) were PCR-amplified from plasmid pFT1 [8] using Pfu polymerase and oligonucleotide primers EGFfw 7-9 (5 -TAGTAGGGATCCATAGAAGGACGATCAGCAGTGGA-TATCGACGAGTGCTCCAAG-3 ), which incorporates a Bam HI site (underlined), factor Xa cleavage site (double underlined) and two spacer amino acids (A, V) upstream of the start of the cbEGF 7 sequence; and EGFrv 7-9 (5 -TAGTAGAAGCTTCTATTAACTGCACACCTTTCCGTC-GCC-3 ), which incorporates a Hind III site (underlined) and two stop codons downstream of the cbEGF 9 sequence. The authenticity of the amplified product was confirmed by DNA sequencing and it was ligated into plasmid expression vector pQ30 (Qiagen) in frame with the N-terminal (His) 6 tag. Recombinant plasmid pEGF 7-9 pQ30 was transformed into E. coli NMR554 [10], which contains the lac repressor on plasmid pRep4.…”

Section: Methodsmentioning

confidence: 99%

“…EGF-like repeats are widely distributed in nature and cbEGFs within extracellular proteins are involved in mediating protein-protein and protein-carbohydrate binding interactions [6]. They are also structurally important in functionally diverse processes, such as maintenance of extracellular matrix architecture, blood coagulation and cholesterol uptake [7].…”

Section: Introductionmentioning

confidence: 99%

Calcium binding activity of the epidermal growth factor-like domains of the apicomplexan microneme protein EtMIC4

Periz

Gill

Knott

et al. 2005

Molecular and Biochemical Parasitology

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Microneme proteins are secreted from apicomplexan parasites during invasion of host cells and they play crucial roles in parasite–host cell adhesion. EtMIC4 is a 240 kDa transmembrane protein from Eimeria tenella that contains 31 tandemly arranged epidermal growth factor (EGF), like repeats within its extracellular domain. The majority of these repeats have calcium binding (cb) consensus sequences. Little is known about cbEGFs in apicomplexan parasites but their presence in microneme proteins suggests that they may contribute to parasite–host interactions. To investigate the potential role of cbEGFs we have expressed and correctly refolded a cbEGF triplet from EtMIC4 (cbEGF7–9) and demonstrated that this triplet binds calcium. Circular dichroism spectroscopic analysis of cbEGF7–9 demonstrates that the molecule undergoes a gradual change in conformation with increasing levels of calcium. In the presence of calcium, the triplet becomes resistant to proteolytic degradation by a variety of proteases, a characteristic feature of cbEGF repeats from higher eukaryotic proteins, such as fibrillin, suggesting that calcium binding induces the formation of a rigid conformation. Moreover, mass spectrometric mapping of the cleavage sites that are protected by calcium shows that these sites are located both close to and distant from the calcium binding sites, indicating that protection is not due to steric hindrance by calcium ions, but rather due to the overall conformation adopted by the triplet in the presence of calcium. Thus, the tandemly-arranged cbEGF repeats within EtMIC4 provide a mechanism whereby, in the calcium-rich extracellular environment, the molecule could adopt a protease-resistant, rigid structure that could favour its interaction with host cell ligands

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The epidermal growth factor–like domains of the human EMR2 receptor mediate cell attachment through chondroitin sulfate glycosaminoglycans

Cited by 195 publications

References 50 publications

N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

EMR1, the human homolog of F4/80, is an eosinophil‐specific receptor

Calcium binding activity of the epidermal growth factor-like domains of the apicomplexan microneme protein EtMIC4

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