title: Tailor-Made Protein Tyrosine Phosphatases: In Vitro Site-Directed Mutagenesis of PTEN and PTPRZ-B.
abstract: In vitro site-directed mutagenesis (SDM) of protein tyrosine phosphatases (PTPs) is a commonly used approach to experimentally analyze PTP functions at the molecular and cellular level and to establish functional correlations with PTP alterations found in human disease. Here, using the tumor-suppressor PTEN and the receptor-type PTPRZ-B (short isoform from PTPRZ1 gene) phosphatases as examples, we provide a brief insight into the utility of specific mutations in the experimental analysis of PTP functions. We describe a standardized, rapid, and simple method of mutagenesis to perform single and multiple amino acid substitutions, as well as deletions of short nucleotide sequences, based on one-step inverse PCR and DpnI restriction enzyme treatment. This method of SDM is generally applicable to any other protein of interest.
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain.
authors: Sandra
authors: S
authors: Luna
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain.
authors: Janire
authors: J
authors: Mingo
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain.
authors: Olaia
authors: O
authors: Aurtenetxe
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain.
authors: Lorena
authors: L
authors: Blanco
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain.
authors: Laura
authors: L
authors: Amo
authors: Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, 6525 GA, Nijmegen, The Netherlands.
authors: Jan
authors: J
authors: Schepens
authors: Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, 6525 GA, Nijmegen, The Netherlands.
authors: Wiljan J
authors: WJ
authors: Hendriks
authors: Biocruces Health Research Institute, Pza Cruces s/n, 48903, Barakaldo, Spain. rpulidomurillo@gmail.com.|IKERBASQUE, Basque Foundation for Science, 48013, Bilbao, Spain. rpulidomurillo@gmail.com.
authors: Rafael
authors: R
authors: Pulido
mesh_terms: D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation; D051059:PTEN Phosphohydrolase / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5 / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; C516895:PTPRZ1 protein, human; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human
keywords: PCR; PTEN; PTPRZ-B; PTPRZ1; Protein tyrosine phosphatase; Site-directed mutagenesis