Introduction of protein kinase recognition sites into proteins: a review of their preparation, advantages, and applications.
Labeled proteins are used in a variety of applications. This review focuses on methods that utilize genetic engineering to introduce protein kinase recognition sites into proteins. Many protein kinase recognition sites can be introduced into proteins and serve as useful tags for a variety of purposes. The introduction of protein kinase recognition sites into proteins can be achieved without modifying the essential structure or function of the proteins. Because proteins modified by these procedures retain their activity after phosphorylation, they can be used in many applications. The phosphorylatable proteins can be labeled easily to high specific activity with radioisotopes ((32)P, (33)P, or (35)S), or the nonradioactive (31)P can be used. The use of these radioisotopes provides a convenient and safe method for radiolabeling proteins. Moreover, the use of the nonradioactive (31)P with protein tyrosine kinase recognition sites permits the tagging of proteins and their detection with the many anti-phosphotyrosine antibodies available. Overall, the procedure represents a convenient, safe, and efficient method to label proteins for a variety of applications.
Pestka
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Department of Molecular Genetics and Microbiology, Robert Wood Johnson Medical School-UMDNJ, 675 Hoes Lane, Piscataway, New Jersey, 08854-5635, USA.
Lin
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L
Wu
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W
Izotova
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L
D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D010766:Phosphorylation; D015202:Protein Engineering / Q000379:methods*; D011494:Protein Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D011494:Protein Kinases