Use of two‐dimensional gel electrophoresis and autoradiography as a tool in cell biology: The example of the thyroid and the liver

Lecocq, Raymond; Lamy, Françoise; Dumont, Jacques Emile

doi:10.1002/elps.1150110303

Cited by 26 publications

(17 citation statements)

References 35 publications

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“…Therefore, we hypothesized that CAPS may promote tumorigenesis and tumor progression by disturbing intracellular Ca 2+ homeostasis. Aprevious study reported that CAPS was phosphorylated and upregulated in response to thyrotropin and the cAMP cascade, and down regulated by TPA and epidermal growth factor (9). CAPS may also contribute to tumorigenesis and tumor progression through crosstalk between cAMP and EGF signals.…”

Section: Discussionmentioning

confidence: 99%

“…A previous study revealed that type-I CAPS may be specific to mammals, type-II CAPS widely exists in metazoan speciesand CAPS 2 is unique to human beings (8). The synthesis and phosphorylation of type-I CAPS are upregulated by thyrotropin and cyclic AMP analogues that can promote cell proliferation and maintain expression of the differentiated thyrocyte phenotype, and are downregulated by 12-O-tetradecanoylphorbol-13-acetate (TPA) and epidermal growth factor, which repress cell differentiation (9). As a member of the EF hand motif family, CAPS contains four EF-hand domains for calcium binding (10).…”

Section: Introductionmentioning

confidence: 99%

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Overexpression of calcyphosine is associated with poor prognosis in esophageal squamous cell carcinoma

Zhu

Yang

et al. 2017

Oncol Lett

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Abstract. Calcyphosine (CAPS), a calcium-binding protein, has been identified as a potential diagnostic and prognostic biomarker in several human carcinomas. However, little is known about CAPS in esophageal squamous cell carcinoma (ESCC). The present study aimed to investigate the expression levels of CAPS in ESCC tissues and evaluate its clinicopathological significance. Reverse transcription-quantitative polymerase chain reaction and immunohistochemical staining were conducted to detect the expression of CAPS in ESCC tissues and adjacent non-cancerous tissues. ESCC samples exhibited higher levels of CAPS mRNA than paired non-cancerous samples (P=0.0015), and the mRNA level of CAPS was positively associated with histological grade (P=0.0013) and tumor invasion depth (P=0.0206). In addition, Kaplan-Meier survival analysis revealed that patients with high CAPS expression experienced significantly shorter 5-year overall survival times than those with low CAPS expression (P=0.0112). Multivariate analysis demonstrated that CAPS protein expression was an independent prognostic biomarker for patients with ESCC. In conclusion, the findings of the present study demonstrated that CAPS may represent a novel diagnostic indicator and an independent prognostic biomarker in ESCC.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Overexpression of calcyphosine is associated with poor prognosis in esophageal squamous cell carcinoma

Zhu

Yang

et al. 2017

Oncol Lett

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“…For immunodetection, proteins were transferred to a nitrocellulose membrane PH 79 (Schleicher & Schiill), for 16 h at 60 V and 4°C as described [25]. The following antibodies were used: mouse anti-c-inyc monoclonal antibody 9.…”

Section: G T C C T G T a T T T Q Q T T T A A T Q T T T T T Q T C C T~mentioning

confidence: 99%

Characterization of a Phosphoprotein whose mRNA is Regulated by the Mitogenic Pathways in Dog Thyroid Cells

Wilkin

Suarez-Huerta

Robaye

et al. 1997

European Journal of Biochemistry

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We have isolated cDNA clones encoding the dog and human forms of a novel protein whose function is still unknown. Sequence analysis indicates that dog clone c5fw protein contains 343 amino acid residues, several potential phosphorylation sites, and two of the 12 conserved subdomains (MI1 and IX) that fold into a common catalytic core structure of the large family of protein kinases. Human clone c S 'shares 95% amino acid identity with its dog counterpart. We have also isolated another human-related clone c53v sharing 70% amino acid identity with the dog sequence. We transiently expressed c-myc epitope-tagged clone c5& protein in COS-7 cells and infected thyrocytes in primary culture with a recombinant adenovirus containing clone c5jw cDNA (adenovirus c5fw). In both experiments, a 46-kDa protein was detected and subsequently more extensively characterized. By two-dimensional gel electrophoresis and V8 protease digestion, we showed that this overexpressed protein is phosphorylated on different sites. Moreover, cells stimulated with thyrotropin or epiderinal growth factor, thyrotropin and fetal calf serum increased the level of clone cSfLL' protein produced after infection by adenovirus containing clone c5fw. The disappearance of this 46-kDa protein after 1 h of puromycin treatment indicates that it is a labile protein. Immunofluorescence and subcellular fractionation analysis have revealed that c -m ytagged clone cSfi was insoluble and localized mainly in the cytoplasm, in the form of granules.

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“…Its synthesis is decreased by epidermal growth factor and phorbol 12-myristate 13-acetate which also trigger cell proliferation but repress expression of differentiation (Lecocq et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

“…Polyclonal antibodies against calcyphosine (Lecocq et al, 1990) have been used as a tool to isolate cDNA encoding calcyphosine (Lefort et al, 1989) …”

Section: Introductionmentioning

confidence: 99%

Rapid purification and identification of calcyphosine, a Ca2+-binding protein phosphorylated by protein kinase A

Lecocq¹,

Lamy²,

Erneux³

et al. 1995

Biochemical Journal

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A method is presented for the rapid purification of dog thyroid calcyphosine, a protein previously identified as a major substrate for cyclic AMP-dependent protein kinase in dog thyroid slices stimulated by thyrotropin [Lecocq, Lamy and Dumont (1979) Eur. J. Biochem. 102, 147-152]. The protein was previously identified as a spot on two-dimensional gels and is now purified in its native form by a procedure involving three chromatographic steps. Homogeneous calcyphosine identified by SDS/PAGE, immunoblotting and peptide sequencing can be obtained within 7 h. As for calmodulin, Ca(2+)-dependent conformational changes can be shown by Ca(2+)-dependent hydrophobic interaction chromatography using phenyl-Sepharose. Unlike calmodulin, calcyphosine is a substrate for protein kinase A.

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Use of two‐dimensional gel electrophoresis and autoradiography as a tool in cell biology: The example of the thyroid and the liver

Abstract: Different applications of two-dimensional gel electrophoresis and the research strategies that this methodology allows, with examples drawn from our own work on thyroid and liver cells, are described.

Cited by 26 publications

References 35 publications

Overexpression of calcyphosine is associated with poor prognosis in esophageal squamous cell carcinoma

Overexpression of calcyphosine is associated with poor prognosis in esophageal squamous cell carcinoma

Characterization of a Phosphoprotein whose mRNA is Regulated by the Mitogenic Pathways in Dog Thyroid Cells

Rapid purification and identification of calcyphosine, a Ca2+-binding protein phosphorylated by protein kinase A

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