Differential expression of the ED sequence-containing form of cellular fibronectin in embryonic and adult human tissues

1987 ◽  
Vol 88 (4) ◽  
pp. 419-430
Author(s):  
T. Vartio ◽  
L. Laitinen ◽  
O. Narvanen ◽  
M. Cutolo ◽  
L.E. Thornell ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Antigenic Determinant ◽  
Solid Phase ◽  
Basement Membranes ◽  
Malignant Tumours ◽  
Embryonic Tissues ◽  
Adult Tissues ◽  
Decidual Cells ◽  
Foetal Kidney ◽  
Cellular Fibronectin

Monoclonal mouse hybridoma antibodies were obtained for secreted cellular fibronectin (cFn) from A8387 fibrosarcoma cells. One of them, 52-DH1 (DH), reacted exclusively with cFns but not with plasma Fns (pFns) in immunoblotting and solid-phase EIA. The DH antibody also recognized thermolysin cFn fragments and beta-galactosidase-Fn fusion protein which contained the ED sequence specific to at least some forms of cFns. On the other hand, the DH antibody failed to recognize a fusion protein that was otherwise identical but lacked the ED sequence. Thus, the antigenic determinant for the DH antibody was located to the ED sequence. The DH antibody was then used to study the expression of ED sequence containing cFn (EcFn). For comparisons, another monoclonal antibody, 52BF12 (BF), recognizing equally well both pFns and cFns, was used. Immunoblotting of pFn fragments indicated that this antibody had the antigenic determinant at or close to the cell-binding site of Fn. EcFn was revealed by the DH antibody in embryonic and adult fibroblasts and in a variety of other cultured normal and malignant human cells. In embryonic tissues EcFn was abundant in developing basement membranes, as shown in foetal kidney and muscle, while in adult tissues it was confined only to endothelia of larger blood vessels. Furthermore, in embryonic tissues the capillaries showed bright EcFn-positivity not found any more in adult tissues. Human plasma contained a small quantity of EcFn, which may hence have an endothelial origin. EcFn was also prominent in the stroma of malignant tumours as well as in reactive benign conditions, such as granulation tissue and decidual cells. The results suggest that EcFn is a form of the protein which may have a particular role in developing and reactive tissues in embryos and adults.

Laminin-521 is a Novel Target of Autoantibodies Associated with Lung Hemorrhage in Anti-GBM Disease

2021 ◽  
pp. ASN.2020101431
Author(s):  
Cong-rong Shen ◽  
Xiao-yu Jia ◽  
Wentian Luo ◽  
Florina Olaru ◽  
Zhao Cui ◽  
...  
Keyword(s):  
Solid Phase ◽  
Pulmonary Hemorrhage ◽  
Basement Membranes ◽  
Glomerular Diseases ◽  
Igg Binding ◽  
Lung Hemorrhage ◽  
Pathogenic Antibodies ◽  
Novel Target ◽  
Circulating Autoantibodies ◽  
Cryptic Epitopes

BackgroundAntiglomerular basement membrane (anti-GBM) disease is characterized by GN and often pulmonary hemorrhage, mediated by autoantibodies that typically recognize cryptic epitopes within α345(IV) collagen—a major component of the glomerular and alveolar basement membranes. Laminin-521 is another major GBM component and a proven target of pathogenic antibodies mediating GN in animal models. Whether laminin-521 is a target of autoimmunity in human anti-GBM disease is not yet known.MethodsA retrospective study of circulating autoantibodies from 101 patients with anti-GBM/Goodpasture’s disease and 85 controls used a solid-phase immunoassay to measure IgG binding to human recombinant laminin-521 with native-like structure and activity.ResultsCirculating IgG autoantibodies binding to laminin-521 were found in about one third of patients with anti-GBM antibody GN, but were not detected in healthy controls or in patients with other glomerular diseases. Autoreactivity toward laminin-521 was significantly more common in patients with anti-GBM GN and lung hemorrhage, compared with those with kidney-limited disease (51.5% versus 23.5%, P=0.005). Antilaminin-521 autoantibodies were predominantly of IgG1 and IgG4 subclasses and significantly associated with lung hemorrhage (P=0.005), hemoptysis (P=0.008), and smoking (P=0.01), although not with proteinuria or serum creatinine at diagnosis.ConclusionsBesides α345(IV) collagen, laminin-521 is another major autoantigen targeted in anti-GBM disease. Autoantibodies to laminin-521 may have the potential to promote lung injury in anti-GBM disease by increasing the total amount of IgG bound to the alveolar basement membranes.

scholarly journals Mouse matriptase-2: identification, characterization and comparative mRNA expression analysis with mouse hepsin in adult and embryonic tissues

2003 ◽  
Vol 373 (3) ◽  
pp. 689-702 ◽  
Author(s):  
John D. HOOPER ◽  
Luisa CAMPAGNOLO ◽  
Goodarz GOODARZI ◽  
Tony N. TRUONG ◽  
Heidi STUHLMANN ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Serine Protease ◽  
Low Density Lipoprotein ◽  
Surface Membrane ◽  
Density Lipoprotein ◽  
Protein Domain ◽  
Complement Protein ◽  
Embryonic Tissues ◽  
Adult Tissues

We report the identification and characterization of mouse matriptase-2 (m-matriptase-2), an 811-amino-acid protein composed of an N-terminal cytoplasmic domain, a membrane-spanning domain, two CUB (complement protein subcomponents C1r/C1s, urchin embryonic growth factor and bone morphogenetic protein 1) domains, three LDLR (low-density-lipoprotein receptor class A) domains and a C-terminal serine-protease domain. All m-matriptase-2 protein domain boundaries corresponded with intron/exon junctions of the encoding gene, which spans approx. 29 kb and comprises 18 exons. Matriptase-2 is highly conserved in human, mouse and rat, with the rat matriptase-2 gene (r-maltriptase-2) predicted to encode transmembrane and soluble isoforms. Western-blot analysis indicated that m-matriptase-2 migrates close to its theoretical molecular mass of 91 kDa, and immunofluorescence analysis was consistent with the proposed surface membrane localization of this protein. Reverse-transcription PCR and in-situ-hybridization analysis indicated that m-matriptase-2 expression overlaps with the distribution of mouse hepsin (m-hepsin, a cell-surface serine protease identified in hepatoma cells) in adult tissues and during embryonic development. In adult tissues both are expressed at highest levels in liver, kidney and uterus. During embryogenesis m-matriptase-2 expression peaked between days 12.5 and 15.5. m-hepsin expression was biphasic, with peaks at day 7.5 to 8.5 and again between days 12.5 and 15.5. In situ hybridization of embryonic tissues indicated abundant expression of both m-matriptase-2 and m-hepsin in the developing liver and at lower levels in developing pharyngo–tympanic tubes. While m-hepsin was detected in the residual embryonic yolk sac and with lower intensity in lung, heart, gastrointestinal tract, developing kidney tubules and epithelium of the oral cavity, m-matriptase-2 was absent in these tissues, but strongly expressed within the nasal cavity by olfactory epithelial cells. Mechanistic insight into the potential role of this new transmembrane serine protease is provided by its novel expression profile in embryonic and adult mouse.

Expression of the heparin-binding cytokines, midkine (MK) and HB-GAM (pleiotrophin) is associated with epithelial-mesenchymal interactions during fetal development and organogenesis

Development ◽  
1995 ◽  
Vol 121 (1) ◽  
pp. 37-51 ◽  
Author(s):  
T.A. Mitsiadis ◽  
M. Salmivirta ◽  
T. Muramatsu ◽  
H. Muramatsu ◽  
H. Rauvala ◽  
...  
Keyword(s):  
Solid Phase ◽  
Biological Effects ◽  
Mesenchymal Cell ◽  
Basement Membranes ◽  
Dependent Manner ◽  
Heparin Binding ◽  
Limb Buds ◽  
New Family ◽  
A Cell

Midkine (MK) and heparin binding-growth associated molecule (HB-GAM or pleiotrophin), constitute a new family of heparin-binding proteins implicated in the regulation of growth and differentiation (T. Muramatsu (1993) Int. J. Dev. Biol. 37, 183–188). We used affinity-purified antibodies against MK and HB-GAM to analyze their distribution during mouse embryonic development. From 9 to 14.5 day post-coitum (dpc), both proteins were detected in central and peripheral nervous systems, facial processes, limb buds, sense organs, respiratory, digestive, urogenital, and skeletal systems. MK and HB-GAM were often localized on the surface of differentiating cells and in basement membranes of organs undergoing epithelial-mesenchymal interactions. The level of MK protein decreased considerably in the 16.5 dpc embryo, whereas HB-GAM staining persisted in many tissues. Our in situ hybridization results revealed a widespread expression of MK transcripts that was not always consistent with the distribution of MK protein in developing tissues. In many epithelio-mesenchymal organs MK and HB-GAM were codistributed with syndecan-1, a cell surface proteoglycan. In limb buds and facial processes, MK, HB-GAM, and syndecan-1 were localized to the apical epithelium and the adjacent proliferating mesenchyme. Both MK and HB-GAM bound syndecan-1 in solid-phase assays in a heparan sulfate-dependent manner. The biological effects of MK and HB-GAM on limb and facial mesenchyme were studied in vitro by application of beads preloaded with the proteins. Neither MK nor HB-GAM stimulated mesenchymal cell proliferation or induced syndecan-1 expression. Taken together these results indicate that MK and HB-GAM may play regulatory roles in differentiation and morphogenesis of the vertebrate embryo, particularly in epithelio-mesenchymal organs, and suggest molecular interactions with syndecan-1.

Tenascin Mr 220,000 isoform expression correlates with corneal cell migration

Development ◽  
1991 ◽  
Vol 112 (2) ◽  
pp. 605-614 ◽  
Author(s):  
A. Kaplony ◽  
D.R. Zimmermann ◽  
R.W. Fischer ◽  
B.A. Imhof ◽  
B.F. Odermatt ◽  
...  
Keyword(s):  
Cell Migration ◽  
Close Correlation ◽  
Basement Membranes ◽  
Embryonic Tissues ◽  
Corneal Fibroblasts ◽  
Sds Page ◽  
Brain Extracts ◽  
Fibronectin Type Iii ◽  
Alternatively Spliced ◽  
Fibronectin Type

The three isoforms of chicken tenascin, an extracellular matrix glycoprotein, are generated by alternatively spliced fibronectin type III domains. The resulting proteins migrate as bands of Mr 220,000 (ten220), Mr 200,000 (ten200) and Mr 190,000 (ten190) on SDS-PAGE. We describe here two monoclonal antibodies, one specific for ten220 (mAb T17) and another that recognizes all isoforms (mAb T16). These were used to examine the differential expression of isoforms during development. Most impressive is the close correlation between ten220 expression and cell migration in the embryonic cornea. Initially (stage 18), ten190/200 can be detected within the corneal epithelium and along the basement membranes of the lens and sclera. Ten220 appears within the primary stroma immediately prior to the invasion by neural-crest-derived cells. This expression is maintained during the subsequent migration of fibroblasts from the conjunctiva into the primary stroma. With the completion of migration and the marked increase in matrix synthesis by corneal fibroblasts, ten220 disappears. Ten190/200 remains in the region adjoining the endothelium, the Bowman's membrane and the adjacent stroma. The cell-migration-associated isoform is isolated from extracts of embryonic tissues as a homohexamer. Low molecular weight forms appeared absent but a new tenascin band of Mr 210,000 could be detected in brain extracts which may be a new isoform. We conclude that the synthesis of tenascin isoforms is under tight developmental control and speculate that a function of the additional domains is to facilitate cell migration.

scholarly journals Solid-phase refolding of poly-lysine tagged fusion protein of hEGF and angiogenin

2002 ◽  
Vol 7 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Sang Joong Park ◽  
Kang Ryu ◽  
Chang Woo Suh ◽  
Young Gyu Chai ◽  
Oh Byung Kwon ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Solid Phase

A STUDY OF ANTIGENS IN ADULT AND EMBRYONIC TISSUES OF THE SLUG DEROCERAS

1965 ◽  
Vol 43 (1) ◽  
pp. 1-12 ◽  
Author(s):  
G. Mattinson
Keyword(s):  
Common Duct ◽  
Blastula Stage ◽  
Stages Of Development ◽  
Agar Gel ◽  
Embryonic Tissues ◽  
Adult Tissues ◽  
Immunofluorescence Studies ◽  
Stage Embryo ◽  
Mixed Stage ◽  
Gel Diffusion

Antigens of embryonic and adult tissues of the slug Deroceras were studied, using the techniques of agar gel diffusion and immunoelectrophoresis. Antibodies were prepared against the tissues by injection of saline homogenates into the ear of a rabbit. Antisera were prepared against whole adult homogenate and against mixed-stage embryo homogenate. The antisera obtained were tested against homogenates of a number of different adult tissues and embryo stages.Twelve or 13 antigens were found in all. Four or five of these antigens were found in embryos only and these antigens were limited to the gastrular and pregastrular stages of development. Two other antigens were found which were specific for adult blood. Five antigens appeared only in embryo and common duct homogenate reactions. One further antigen was found in three adult tissues and in embryos from the blastula stage onward. Immunofluorescence studies showed that the antigens were located partially or wholly in the cellular tissues of the embryos and not in the surrounding membranes. The antigens were located in the cytoplasm at all stages, and in the nuclei and nucleoli of one particular type of cell in the gastrular stages.

Covalent immobilization and solid-phase refolding of enterokinase for fusion protein cleavage

2005 ◽  
Vol 40 (5) ◽  
pp. 1755-1762 ◽  
Author(s):  
Chang Woo Suh ◽  
Sin Hye Park ◽  
Seung Gook Park ◽  
Eun Kyu Lee
Keyword(s):  
Fusion Protein ◽  
Solid Phase ◽  
Covalent Immobilization ◽  
Protein Cleavage ◽  
Fusion Protein Cleavage

Solid-phase synthesis and application of double-fluorescent-labeled lipopeptides, containing a CTL-epitope from the measles fusion protein

1999 ◽  
Vol 54 (5) ◽  
pp. 436-443 ◽  
Author(s):  
P. Hoogerhout ◽  
K.J. Stittelaar ◽  
H.F. Brugghe ◽  
J. A. M. Timmermans ◽  
G.J. Ten Hove ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Phase Synthesis ◽  
Ctl Epitope

SPECIFIC INHIBITION OF DIFFERENTIATION IN THE FROG EMBRYO BY CELL-FREE HOMOGENATES OF ADULT TISSUES

1959 ◽  
Vol 37 (2) ◽  
pp. 129-131 ◽  
Author(s):  
R. B. Clarke ◽  
D. J. McCallion
Keyword(s):  
Adult Brain ◽  
Specific Inhibition ◽  
Frog Embryo ◽  
Embryonic Tissues ◽  
Adult Tissues ◽  
Inhibition Of Differentiation

The hypothesis that adult tissues produce substances which may specifically inhibit the differentiation of like embryonic tissues and organs has been tested by Rose (3) by rearing frog embryos in the presence of living fragments of adult organs. In the present investigation frog embryos were reared in media containing cell-free homogenates of adult brain or heart. The homogenates specifically suppressed or inhibited the development of these organs in about 10–15% of the embryos tested.

scholarly journals The C-Terminal Portion of the Tail Fiber Protein of Bacteriophage Lambda Is Responsible for Binding to LamB, Its Receptor at the Surface of Escherichia coli K-12

2000 ◽  
Vol 182 (2) ◽  
pp. 508-512 ◽  
Author(s):  
Jiang Wang ◽  
Maurice Hofnung ◽  
Alain Charbit
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Solid Phase ◽  
Cell Surface Receptor ◽  
Tail Fiber ◽  
Tail Fiber Protein ◽  
Intact Cells ◽  
Fiber Protein ◽  
Surface Receptor ◽  
K 12

ABSTRACT Bacteriophage λ adsorbs to its Escherichia coli K-12 host by interacting with LamB, its cell-surface receptor. We fused C-terminal portions of J, the tail fiber protein of λ, to maltose-binding protein. Solid-phase binding assays demonstrated that a purified fusion protein comprising only the last 249 residues of J could bind to LamB trimers and inhibited recognition by anti-LamB antibodies. Electron microscopy further demonstrated that the fusion protein could also bind to LamB at the surface of intact cells. This interaction prevented λ adsorption but affected only partially maltose uptake.

Sign in / Sign up

Export Citation Format

Share Document