Ependymins from the cerebrospinal fluid of salmonid fish: gene structure and molecular characterization

Müller-Schmid, Angelika; Rinder, Heinz; Lottspeich, Friedrich; Gertzen, Eva-Maria; Hoffmann, Werner

doi:10.1016/0378-1119(92)90188-u

Cited by 35 publications

(26 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Since the Ca2+-binding capacity is still present at a very low level even after digestion with peptide N-glycosidase F (Fig. 2B), certain acidic clustcrs at positions 112-114 and 181-184 in the protein backbone of rainbow trout ependymins (Muller-Schmid et al, 1992) may also contribute slightly to this affinity.…”

Section: Ca" Bindingmentioning

confidence: 99%

“…The ependymin antisera used in this study were PCa-1 against the C-terminal region of goldfish ependymins (Konigstorfer et al, 1989b) and EOm-1 against purified ependymin A from rainbow trout (Muller-Schmid et al, 1992). column using buffer A (50mM Tris, pH 7.5, 2.5 mM CaCI,) and buffer B (50 mM Tris, pH 7.5, 2.5 mM CaCl,, 1 M NaCl).…”

Section: Analysis Of Ependyminsmentioning

confidence: 99%

“…Ependymins, erroneously named after their first immunohistochemical localization in goldfish ependymal zones (Shashoua, 1985), are secretory glycoproteins of the endomeninx (Konigslorfcr et al, 1990;Sterrer et al, 1990;Rinder et al, 1992;Schwarz et al, 1993). Soluble forms represent the predominant constituents of the cerebrospinal fluid (CSF) of some orders of teleost fish (HolTmann, 1992) and a bound form is associated with the extracellular matrix (Thormodsson et al, 1992a;Schwarz et al, 1993).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Calcium binding to sialic acids and its effect on the conformation of ependymins

Ganß

Hoffmann

1993

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

Soluble ependymins form the predominant protein constituents in the cerebrospinal fluid from many orders of teleost fish. Furthermore, these glycoproteins also exist in a bound form associated with the extracellular matrix. Ependymins are synthesized in meningeal fibroblasts. In goldfish, their synthesis is increased during the regeneration of the optic nerve and they share several characteristics with molecules involved in cell contact phenomena. In this study, we show by a calcium overlay technique that ependymins from goldfish and rainbow trout are able to bind 45Ca2+. However, nearly all of this Ca2+‐binding capacity is lost after digestion with sialidase. Furthermore, circular‐dichroism spectra from FPLC‐purified rainbow trout ependymins have been recorded in the presence and absence of Ca2+. Below 250 nm, the CD spectrum showed a characteristic minimum of ellipticity at 217 nm typical of β structures. This signal is independent of the Ca2+ concentration. In contrast, the complex signal at 250–310 nm mainly decreased with increasing Ca2+ concentration indicating changes in the environment of aromatic side chains.

show abstract

Section: Ca" Bindingmentioning

confidence: 99%

Section: Analysis Of Ependyminsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Calcium binding to sialic acids and its effect on the conformation of ependymins

Ganß

Hoffmann

1993

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…As a control, the blot was rehybridized with the 32P-labeled insert of pGAD-28 (31) encoding chicken glyceraldehyde-3-phosphate dehydrogenase. Southern blot analysis, with 25 ,ug of restriction endonuclease-digested human genomic DNA used per lane, was done according to Muller-Schmid et al (32).…”

mentioning

confidence: 99%

hP1.B, a human P-domain peptide homologous with rat intestinal trefoil factor, is expressed also in the ulcer-associated cell lineage and the uterus.

Hauser

Poulsom

Chinery

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

149

140

View full text Add to dashboard Cite

The six-cysteine P-domain motif forms the basic repeat unit of a growing family of mucin-associated peptides. A precursor for a human secretory polypeptide has been discovered by molecular cloning and deduced to have a single P-domain, termed hP1.B. The pre-pro-peptide has 67% amino acid identity with rat intestinal trefoil factor. We find, using the techniques of RNA analysis and in situ hybridization, that this P-domain peptide is expressed in the human gastrointestinal tract, where a number of pathological conditions affect its expression, and surprisingly find it is expressed in the uterus also. In the intestine, hP1.B is expressed by goblet cells, but in Crohn disease this peptide is synthesized and secreted additionally by the ulcer-associated cell lineage that is known to secrete two other trefoil peptides, pS2 and spasmolytic polypeptide (hSP). In the stomach, hP1.B mRNA is relatively scarce but is more abundant in foci of intestinal metaplasia and near to ulceration. Mucin-rich epithelial cells in hyperplastic polyps of the colon also express this peptide. The discovery of this P-domain peptide and its expression in association with mucins support the hypothesis that P-domains with mucins may subserve related functions in the maintenance and repair of mucosal function.

show abstract

“…EPNs in general contain endoplasmic reticulum-targeting signal sequences at the N-terminus, which are likely cleaved by signal peptidases and further processed for secretion [9,11]. The sequences also hold four cysteine residues that may form either intramolecular disulfide crosslinking for stabilizing protein conformation or intermolecular crosslinking for mediating dimeric interaction ( Figure 1).…”

Section: Introductionmentioning

confidence: 99%

Over-Expression, Secondary Structure Characterization, and Preliminary X-ray Crystallographic Analysis of Xenopus tropicalis Ependymin

2018

View full text Add to dashboard Cite

Abstract:The gene encoding frog (Xenopus tropicalis) ependymin without the signaling sequence was gene-synthesized, and the protein successfully over-expressed in~mg quantities adequate for crystallization using insect cell expression. Circular dichroism (CD) analysis of the protein purified with >95% homogeneity indicated that ependymin contains both α-helix and β-strand among the secondary structure elements. The protein was further crystallized using polyethylene glycol 8000 as the precipitating reagent, and X-ray diffraction data were collected to 2.7 Å resolution under cryo-condition at a synchrotron facility. The crystal belongs to a hexagonal space group P6 1 22 (or P6 5 22) having unit cell parameters of a = b = 61.05 Å, c = 234.33 Å. Matthews coefficient analysis indicated a crystal volume per protein mass (V M ) of 2.76 Å 3 Da −1 and 55.4% solvent content in the crystal when the calculated molecular mass of the protein only was used. However, the apparent SDS-PAGE molecular mass of~33 kDa (likely resulting from N-glycosylation) suggested V M of 1.90 Å 3 Da −1 and 35.4% solvent content instead. In both cases, the asymmetric unit of the crystal likely contains only one subunit of the protein.

show abstract

Ependymins from the cerebrospinal fluid of salmonid fish: gene structure and molecular characterization

Cited by 35 publications

References 24 publications

Calcium binding to sialic acids and its effect on the conformation of ependymins

Calcium binding to sialic acids and its effect on the conformation of ependymins

hP1.B, a human P-domain peptide homologous with rat intestinal trefoil factor, is expressed also in the ulcer-associated cell lineage and the uterus.

Over-Expression, Secondary Structure Characterization, and Preliminary X-ray Crystallographic Analysis of Xenopus tropicalis Ependymin

Contact Info

Product

Resources

About