What question did this study set out to answer?

To develop an enzymatic approach using recombinant lambda protein phosphatase for high-throughput detection and mapping of phosphoproteins on two-dimensional polyacrylamide gels.

September 1, 2002PROTEOMICS

Mapping of phosphorylated proteins on two-dimensional polyacrylamide gels using protein phosphatase

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To develop an enzymatic approach using recombinant lambda protein phosphatase for high-throughput detection and mapping of phosphoproteins on two-dimensional polyacrylamide gels.
Divided cultured rat skin fibroblast proteins into two aliquots, treating one with recombinant lambda protein phosphatase while keeping the other untreated.
Separated both aliquots via two-dimensional polyacrylamide gel electrophoresis and mapped phosphoproteins by tracking charge-dependent spot shifts toward basic positions.
Characterized shifted phosphoprotein spots and detachment-induced phosphorylation changes using mass spectrometry.
Approximately 5% of detectable proteins in cultured rat skin fibroblasts were determined to be phosphorylated based on mobility shifts toward more basic positions.
Mass spectrometry successfully identified 14 distinct phosphoproteins, including proteasome component C8 and small glutamine-rich tetratricopeptide repeat-containing protein.
Phosphorylation of actin-modulating proteins destrin and cofilin decreased significantly following chemical or enzymatic cell detachment from culture dishes.

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Implication

Experimental study demonstrates high-throughput phosphoprotein mapping via phosphatase treatment in rat fibroblasts, indicating a broadly applicable proteomic profiling method.

Cite This Study

Yamagata et al. (2002) studied this question.

synapsesocial.com/papers/6a761711d65efbf82c8c54a4 https://doi.org/10.1002/1615-9861(200209)2:9<1267::aid-prot1267>3.0.co;2-r

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Implication

Experimental study demonstrates high-throughput phosphoprotein mapping via phosphatase treatment in rat fibroblasts, indicating a broadly applicable proteomic profiling method.

Mapping of phosphorylated proteins on two-dimensional polyacrylamide gels using protein phosphatase

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Discussion

Implication

Cite This Study

Mapping of phosphorylated proteins on two-dimensional polyacrylamide gels using protein phosphatase

Key Points

Discussion

Implication

Cite This Study

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