TY - CHAP
T1 - Chapter 1 Protein Phosphorylation by Semisynthesis. From Paper to Practice
AU - Szewczuk, Lawrence M.
AU - Tarrant, Mary Katherine
AU - Cole, Philip A.
N1 - Funding Information:
We are grateful for the insightful comments of many of our colleagues, most of whose names are noted in the references. We thank the National Institutes of Health for financial support.
PY - 2009
Y1 - 2009
N2 - Deconvolution of specific phosphorylation events can be complicated by the reversibility of modification. Protein semisynthesis with phosphonate analogues offers an attractive approach to functional analysis of signaling pathways. In this technique, N- and C-terminal synthetic peptides containing nonhydrolyzable phosphonates at target residues can be ligated to recombinant proteins of interest. The resultant semisynthetic proteins contain site specific, stoichiometric phosphonate modifications and are completely resistant to phosphatases. Control of stoichiometry, specificity, and reversibility allows for complex signaling systems to be broken down into individual events and discretely examined. This chapter outlines the general methods and considerations for designing and carrying out phosphoprotein semisynthetic projects.
AB - Deconvolution of specific phosphorylation events can be complicated by the reversibility of modification. Protein semisynthesis with phosphonate analogues offers an attractive approach to functional analysis of signaling pathways. In this technique, N- and C-terminal synthetic peptides containing nonhydrolyzable phosphonates at target residues can be ligated to recombinant proteins of interest. The resultant semisynthetic proteins contain site specific, stoichiometric phosphonate modifications and are completely resistant to phosphatases. Control of stoichiometry, specificity, and reversibility allows for complex signaling systems to be broken down into individual events and discretely examined. This chapter outlines the general methods and considerations for designing and carrying out phosphoprotein semisynthetic projects.
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U2 - 10.1016/S0076-6879(09)62001-2
DO - 10.1016/S0076-6879(09)62001-2
M3 - Chapter
C2 - 19632467
AN - SCOPUS:67650725201
SN - 9780123743107
T3 - Methods in Enzymology
SP - 1
EP - 24
BT - Methods in Enzymology
A2 - Muir, Tom
A2 - Abelson, John
ER -