[Identification of interaction and interaction domains between neuroglobin and Na(+), K(+)-ATPase beta2 subunit].
The pre-transformed human fetal brain cDNA library was used to screen the protein interacting with neuroglobin by using yeast two hybrid system III from ClonTech Inc. The protein encoded by one of the clones interacting with neuroglobin (NGB) was confirmed to be the C terminus of the Na(+), K(+)-ATPase beta2 subunit (NKA1b2) based on amino acid sequences. Then the full-length coding region cDNA sequence of NKA1b2 was obtained from human fetal brain cDNA library by PCR. A set of experiments were designed to test the interaction between NGB and NKA1b2. Interaction between NGB and NKA1b2 was confirmed by binding assay in vitro. Furthermore, the interaction was also proved by co-immunoprecipitation test in vivo. Moreover, the structure integrity of neuroglobin was found to be essential for the interaction between NGB and NKA1b2 by yeast two hybrid method with a series of neuroglobin truncated mutants.
Xu
Wen-Lin
WL
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Wang
Chun-Li
CL
Liao
Zhi-Yong
ZY
Zhang
Yong-Liang
YL
Yu
Li-Hong
LH
Meng
Fan-Wei
FW
Wang
Xin-Xing
XX
Meng
Fan-Wei
FW
Yin
Zhao-Yun
ZY
Qian
Ling-Jia
LJ
Zhang
Cheng-Gang
CG
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005914:Globins / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D000078787:Neuroglobin; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D005914:Globins; D005982:Glutathione Transferase; D000254:Sodium-Potassium-Exchanging ATPase