TY - JOUR
T1 - The conformational polymorphism of the green fluorescent protein
AU - Tan, Haidong
AU - Li, Yueguang
AU - Chen, Ling
AU - Kudoh, Takayuki
AU - Kasai, Tomonari
AU - Seno, Masaharu
N1 - Funding Information:
ACKNOWLEDGMENTS We appreciate Ms. Mami Asakura for her excellent technical assistance. We also thank the anonymous reviewer for his or her insightful suggestions. The financial support of the National Basic Research Program of China (973 Program) (no. 2007CB707802) is gratefully acknowledged.
PY - 2012/2
Y1 - 2012/2
N2 - 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.
AB - 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.
KW - MALDI-TOF MS
KW - circular dichroism spectra
KW - conformational polymorphism
KW - glutathione-S-transferase
KW - green fluorescent protein
KW - native polyacrylamide gel electrophoresis
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U2 - 10.1134/S0026893311060045
DO - 10.1134/S0026893311060045
M3 - Article
AN - SCOPUS:84863159530
SN - 0026-8933
VL - 46
SP - 142
EP - 148
JO - Molecular Biology
JF - Molecular Biology
IS - 1
ER -