| Chapter title |
Tailor-Made Protein Tyrosine Phosphatases: In Vitro Site-Directed Mutagenesis of PTEN and PTPRZ-B
|
|---|---|
| Chapter number | 5 |
| Book title |
Protein Tyrosine Phosphatases
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3746-2_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3744-8, 978-1-4939-3746-2
|
| Authors |
Sandra Luna, Janire Mingo, Olaia Aurtenetxe, Lorena Blanco, Laura Amo, Jan Schepens, Wiljan J. Hendriks, Rafael Pulido |
| 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 7 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 2 | 29% |
| Student > Ph. D. Student | 2 | 29% |
| Unspecified | 1 | 14% |
| Student > Bachelor | 1 | 14% |
| Student > Master | 1 | 14% |
| Other | 0 | 0% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 43% |
| Unspecified | 1 | 14% |
| Agricultural and Biological Sciences | 1 | 14% |
| Medicine and Dentistry | 1 | 14% |
| Unknown | 1 | 14% |