Development of Specific Protein Labeling Method by Using Small Molecular Probes

作者：Ojida Akio^*

来源：Journal of the Pharmaceutical Society of Japan, 2012, 132(2): 195-199.

DOI：10.1248/yakushi.132.195

• protein labeling
• covalent modification
• molecular probe
• fluorescence
• bioimaging
• G-protein coupled receptor

Selective protein labeling with a small molecular probe is a versatile method for elucidating protein functions in living cells. In this review, we report a new protein labeling method that enables selective covalent modification of a tag-fused protein with small functional molecules. This method utilizes the specific interaction and rapid reaction between a short peptide tag and a molecular probe, which comprises the cysteine-containing short CA6D4x2 tag (CAAAAAA-DDDDGDDDD) and a tetranuclear Zn (II)-DpaTyr probe containing a reactive a-chloroacetyl moiety. This labeling system, so-called reactive tag system, was successfully applied to the fluorescence imaging of tag-fused GPCRs such as bradykinin receptor (B2R) and acetylcholine receptor (m 1 AchR) expressing on HEK293 cells. The utility of this labeling method was demonstrated in the function analyses of GPCRs, such as fluorescence visualization of the stimuli-responsive internalization of GPCRs and pH change in endosomes containing the internalized GPCRs.

• 出版日期2012-2