PubMed:22816877
Annnotations
{"target":"https://pubannotation.org/docs/sourcedb/PubMed/sourceid/22816877","sourcedb":"PubMed","sourceid":"22816877","source_url":"https://www.ncbi.nlm.nih.gov/pubmed/22816877","text":"Control of a tyrosyl radical mediated protein cross-linking reaction by electrostatic interaction.\nHerein, we demonstrate the control of protein heteroconjugation via a tyrosyl coupling reaction by using electrostatic interaction. Aspartic acid and arginine were introduced to a tyrosine containing peptide tag (Y-tag) to provide electrostatic charge. Designed negatively or positively charged Y-tags were tethered to the C-terminus of Escherichia coli alkaline phosphatase (BAP) and streptavidin (SA), and these model proteins were subjected to horseradish peroxidase (HRP) treatment. The negatively charged Y-tags showed low reactivity due to repulsive interactions between the Y-tags with the negatively charged BAP and SA. In contrast, the positively charged Y-tags showed high reactivity, indicating that the electrostatic interaction between Y-tags and proteins significantly affects the tyrosyl radical mediated protein cross-linking. From the heteroconjugation reaction of BAP and SA, the SA with the positively charged Y-tags exhibited favorable cross-linking toward negatively charged BAP, and the BAP-SA conjugates prepared from BAP with GY-tag (GGGGY) and SA with RYR-tag (RRYRR) had the best performance on a biotin-coated microplate. Encompassing the reactive tyrosine residue with arginine residues reduced the reactivity against HRP, enabling the modulation of cross-linking reaction rates with BAP-GY. Thus, by introducing a proper electrostatic interaction to Y-tags, it is possible to kinetically control the heteroconjugation behavior of proteins, thereby maximizing the functions of protein heteroconjugates.","tracks":[{"project":"Allie","denotations":[{"id":"SS1_22816877_3_0","span":{"begin":484,"end":496},"obj":"expanded"},{"id":"SS2_22816877_3_0","span":{"begin":498,"end":500},"obj":"abbr"},{"id":"SS1_22816877_3_1","span":{"begin":546,"end":568},"obj":"expanded"},{"id":"SS2_22816877_3_1","span":{"begin":570,"end":573},"obj":"abbr"}],"relations":[{"id":"AE1_22816877_3_0","pred":"abbreviatedTo","subj":"SS1_22816877_3_0","obj":"SS2_22816877_3_0"},{"id":"AE1_22816877_3_1","pred":"abbreviatedTo","subj":"SS1_22816877_3_1","obj":"SS2_22816877_3_1"}],"attributes":[{"subj":"SS1_22816877_3_0","pred":"source","obj":"Allie"},{"subj":"SS2_22816877_3_0","pred":"source","obj":"Allie"},{"subj":"SS1_22816877_3_1","pred":"source","obj":"Allie"},{"subj":"SS2_22816877_3_1","pred":"source","obj":"Allie"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"Allie","color":"#93d2ec","default":true}]}]}}