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http://purl.uniprot.org/citations/17563729http://www.w3.org/1999/02/22-rdf-syntax-ns#typehttp://purl.uniprot.org/core/Journal_Citation
http://purl.uniprot.org/citations/17563729http://www.w3.org/2000/01/rdf-schema#comment"

Purpose

The current studies were conducted to determine whether the protein tyrosine phosphatase, PTP1B, plays a role in regulating epidermal growth factor receptor (EGFR) Tyr992 phosphorylation and cell cycle entry in rat corneal endothelial cells.

Methods

Corneas were obtained from male Sprague-Dawley rats. PTP1B mRNA and protein expression were compared in confluent and subconfluent cells by RT-PCR and western blots. Immunocytochemistry was used to determine the subcellular localization of both PTP1B and EGFR following epidermal growth factor (EGF) stimulation. Western blots were used to analyze the time-dependent effect of EGF on phosphorylation of EGFR Tyr992 plus or minus CinnGEL 2Me, an inhibitor of PTP1B activity. The effect of PTP1B inhibition on cell cycle entry was determined by calculating the percent of Ki67-positive cells following EGF treatment.

Results

PTP1B mRNA expression was similar in confluent and subconfluent cells, but PTP1B protein was expressed at 3 fold higher levels in subconfluent cells. Positive staining for PTP1B was localized in vesicular structures below the plasma membrane. EGFR staining was located at cell-cell borders in untreated endothelium, but was mainly cytoplasmic by 15 min after EGF treatment. In control cultures, phosphorylation of EGFR Tyr992 peaked by 5 min following EGF stimulation and rapidly decreased to basal levels by 30 min. In cultures pretreated with CinnGEL 2Me, Tyr992 phosphorylation peaked 2 min following EGF addition and was consistently sustained at a higher level than controls until 60 min after treatment. By 18 h following EGF treatment, cultures pretreated with CinnGEL 2Me exhibited a 1.7 fold increase in the number of Ki67-positive cells compared with control cultures.

Conclusions

Comparison of PTP1B mRNA and protein levels indicates that PTP1B expression is regulated mainly at the protein level and is higher in subconfluent cells. PTP1B was located in vesicles below the plasma membrane. The fact that EGFR is internalized in response to EGF stimulation suggests that it could interact with and be regulated by PTP1B. The ability of PTP1B inhibitor to sustain EGFR Tyr992 phosphorylation and increase the number of Ki67-positive cells indicates that PTP1B plays a role in the negative regulation of EGF-induced signaling and helps suppress cell cycle entry."xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/author"Harris D.L."xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/author"Joyce N.C."xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/date"2007"xsd:gYear
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/name"Mol Vis"xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/pages"785-796"xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/title"Protein tyrosine phosphatase, PTP1B, expression and activity in rat corneal endothelial cells."xsd:string
http://purl.uniprot.org/citations/17563729http://purl.uniprot.org/core/volume"13"xsd:string
http://purl.uniprot.org/citations/17563729http://www.w3.org/2004/02/skos/core#exactMatchhttp://purl.uniprot.org/pubmed/17563729
http://purl.uniprot.org/citations/17563729http://xmlns.com/foaf/0.1/primaryTopicOfhttps://pubmed.ncbi.nlm.nih.gov/17563729
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http://purl.uniprot.org/uniprot/P20417http://purl.uniprot.org/core/mappedCitationhttp://purl.uniprot.org/citations/17563729
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