Helena Stabile, Stefania Mitola, Emanuela Moroni, Mirella Belleri, Stefania Nicoli, Daniela Coltrini, Francesco Peri, Antonello Pessi, Laura Orsatti, Fabio Talamo, Vincent Castronovo, David Waltregny, Franco Cotelli, Domenico Ribatti, Marco Presta; Bone morphogenic protein antagonist Drm/gremlin is a novel proangiogenic factor. (B) Negative control in which the primary antibody was omitted. Bovine aortic endothelial (BAE) cells and normal subcutaneous microvascular endothelial (SIE) cells20  (both provided by A. Vecchi, Istituto Mario Negri, Milan, Italy) were cultured in DMEM supplemented with 10% heat-inactivated donor calf serum. Nano-ESI-MS/MS analysis also shows that purified Drm undergoes posttranslational modifications, including maturation, glycosylation, and phosphorylation, in keeping with previous observations in Drm-transfected COS cells.24  Indeed, purified Drm lacks the leader sequence for secretion (amino acid sequence starting from Lys-25) and carries one N-glycosylation at Asn-42 and one phosphorylation at Ser-77 (Figure 1F). After 4 days, blood vessels converging versus the implant were counted. After a PBS wash, cells were washed twice with 2.0 M NaCl in 20 mM HEPES buffer (pH 7.5) to elute 125I-rDrm bound to low-affinity sites. Ten samples of formalin-fixed and paraffin-embedded human lung cancers (5 adenocarcinomas and 5 squamous-cell carcinomas) were obtained from L. de Leval (Department of Pathology, Liège University Hospital, Belgium). The algorithm is described in the ISO 3309 standard. Alterations of blood vessel development by endothelial cells overexpressing fibroblast growth factor-2. Figure 4 Schematic representation of the gremlin 2 syntenic region in pigs. The data presented in this section are a quality-filtered subset of binary interactions automatically derived from the IntAct database. Proangiogenic activity of rDrm. Accordingly, a potent angiogenic response was observed in chick embryo CAMs implanted with Drm-transfectants when compared with mock-transfected cells (Figure S1; Table 1). Data are expressed as mean ± SD. (A) Two-dimensional electrophoresis of control and rDrm-treated SIE-cell extracts decorated with anti–phospho-Tyr antibody. The version number for both the entry and the canonical sequence are also displayed.

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This subsection of the 'Entry information' section indicates whether the entry has been manually annotated and reviewed by UniProtKB curators or not, in other words, if the entry belongs to the Swiss-Prot section of UniProtKB (reviewed) or to the computer-annotated TrEMBL section (unreviewed).

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, The European Molecular Biology Laboratory, State Secretariat for Education, Research and Innovation, transmembrane receptor protein tyrosine kinase activator activity, vascular endothelial growth factor receptor 2 binding, cell migration involved in sprouting angiogenesis, mesenchymal to epithelial transition involved in metanephros morphogenesis, negative regulation of BMP signaling pathway, negative regulation of bone mineralization, negative regulation of bone mineralization involved in bone maturation, negative regulation of bone trabecula formation, negative regulation of canonical Wnt signaling pathway, negative regulation of chondrocyte differentiation, negative regulation of monocyte chemotaxis, negative regulation of osteoblast differentiation, negative regulation of osteoblast proliferation, negative regulation of osteoclast proliferation, negative regulation of pathway-restricted SMAD protein phosphorylation, negative regulation of transcription, DNA-templated, positive regulation of branching involved in ureteric bud morphogenesis, positive regulation of cardiac muscle cell differentiation, positive regulation of cell population proliferation, positive regulation of NF-kappaB transcription factor activity, positive regulation of NIK/NF-kappaB signaling, positive regulation of peptidyl-tyrosine autophosphorylation, positive regulation of receptor internalization, positive regulation of signaling receptor activity, positive regulation of telomerase activity, positive regulation of transcription by RNA polymerase II, positive regulation of transcription from RNA polymerase II promoter involved in myocardial precursor cell differentiation, regulation of epithelial to mesenchymal transition, sequestering of BMP from receptor via BMP binding, Polyposis syndrome, mixed hereditary 1 (HMPS1). Cancer cells and endothelial cells of each specimen were scored for Drm/gremlin immunoreactivity on an arbitrary scale: − indicates negative; +, weak staining; ++, strong staining. Formation of radially growing cell sprouts was observed during the next 24 to 72 hours. RT-PCR analysis of total RNA from these lesions using specie-specific primers confirmed the murine stromal origin of Drm/gremlin and the lack of human Drm/gremlin transcripts (Figure 6). No or very weak immunoreactivity is detected in nonneoplastic lung tissue adjacent to the tumor (D). CAMs were photographed after 4 days. Highly vascularized tumor xenografts generated by subcutaneous injection in nude mice of human adenocarcinoma HEC-1-B–derived cells26  showed a strong Drm immunoractivity in the tumor stroma, whereas tumor parenchyma was negative (Figure 6A-B). Our observations point to a novel, previously unrecognized capacity of Drm/gremlin to interact directly with target endothelial cells and to modulate angiogenesis. designed the research; H.S., S.M., E.M., M.B., S.N., D.C., L.O., V.C., D.W., and D.R. The information is extracted from the scientific literature and diseases that are also described in the OMIM database are represented with a controlled vocabulary in the following way:

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This section describes post-translational modifications (PTMs) and/or processing events.

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This subsection of the 'PTM / Processing' section denotes the presence of an N-terminal signal peptide.

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This subsection of the 'PTM / Processing' section describes the extent of a polypeptide chain in the mature protein following processing or proteolytic cleavage.

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This subsection of the PTM / Processing section specifies the position and type of each covalently attached glycan group (mono-, di-, or polysaccharide).

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This subsection of the PTM / Processing":/help/ptm_processing_section section describes the positions of cysteine residues participating in disulfide bonds.

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Manually validated information inferred from a combination of experimental and computational evidence.

Migrated cells at the bottom surface of the filter were stained (Diff-Quick; DADE Behring, Marburg, Germany) and counted at × 250 magnification (5 fields/sample in triplicate) using a Dialux 20 EB microscope (Leitz, Wetzlar, Germany) equipped with an NPl 25×/0.50 NA objective. Squamous tumor cells are weakly immunoreactive (A). It is updated at every UniProt release.

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This section provides information on the tertiary and secondary structure of a protein.

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This subsection of the 'Structure' section is used to indicate the positions of experimentally determined beta strands within the protein sequence.

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This subsection of the 'Structure' section is used to indicate the positions of experimentally determined helical regions within the protein sequence.

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This subsection of the 'Structure' section is used to indicate the positions of experimentally determined hydrogen-bonded turns within the protein sequence.