Thymidine kinase 1 (TK1) is a cytosolic enzyme essential for thymidine phosphorylation during DNA synthesis. Phosphorylation at Ser13 occurs during mitosis and is mediated by cyclin-dependent kinases (CDKs), particularly CDK1 and CDK2 . Key findings include:
Functional Impact: Mutation of Ser13 to alanine abolishes mitotic phosphorylation without altering TK1’s catalytic activity .
Regulatory Role: Phosphorylation correlates with cell cycle progression, peaking during G2/M phase, and modulates TK1’s interaction with substrates like [18F]FLT, a radiopharmaceutical used in cancer imaging .
Phospho-TK1 (Ser13) antibodies are typically rabbit polyclonal or monoclonal IgG raised against synthetic phosphopeptides.
Western Blot: Distinct bands at ~25 kDa in HCT116, HeLa, and Jurkat cell lysates .
IHC: Strong nuclear/cytoplasmic staining in mouse kidney and testis tissues .
Functional Assays: Reduced [18F]FLT cellular retention in cells with Ser13 mutations .
Mitotic Regulation: Ser13 phosphorylation is a biomarker for mitotic entry, detectable in synchronized cells .
Drug Response: Chemotherapeutic agents like roscovitine (CDK inhibitor) reduce phosphorylation, correlating with G2/M arrest .
Imaging: Phospho-TK1 status influences [18F]FLT uptake in tumors, aiding PET imaging sensitivity .
Prognostic Value: Overexpression of phosphorylated TK1 is linked to aggressive tumors (e.g., melanoma, leukemia) .
Sample Prep: Use phosphatase inhibitors to preserve phosphorylation status .
Controls: Include non-phosphorylated TK1 and Ser13Ala mutants to confirm specificity .
Limitations: Not validated for flow cytometry or diagnostic use .
Current research focuses on: