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Vol 46(2012) N 1 p. 142-148; Haidong Tan1, Yueguang Li2, Ling Chen3, Takayuki Kudoh3, Tomonari Kasai3, Masaharu Seno3* The Сonformational Polymorphism of the Green Fluorescent Protein 1Dalian Institute of Chemical Physics, Dalian, 116023, China2Department of General Surgery, Tianjin 4th Centre Hospital, Tianjin, 300140, China 3Department of Medical and Bioengineering Science, Graduate School of Natural Science and Technology, Okayama University, Okayama, 7008530, Japan *mseno@cc.okayama-u.ac.jp Received - 2010-11-09; Accepted - 2011-01-13 Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. However, its state in liquid and its effect on other proteins are still unclear. The conformational polymorphisms of glutathione-S-transferase-green fluorescent protein (GST-GFPuv), GFPuv and GST were analyzed by native polyacrylamide gel, indicating that GST was in many different states while GFPuv and GST-GFPuv were only in four and two slightly different states. Four different circular dichroism spectra were obtained from the GFPuv polymorphisms. The single molecular behavior of GST-GFPuv and GFPuv was also characterized by MALDI-TOF MS. Thus, we demonstrated that: (1) there might be four different structural polymorphisms for the native GFPuv; (2) GFPuv could reduce its partner's polymorphism as a fusion protein. Although GFPuv had many merits as a reporter, its unreliability was found in the study. green fluorescent protein, conformational polymorphism, glutathione-S-transferase, native polyacrylamide gel electrophoresis, MALDI-TOF MS, circular dichroism spectra |
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