title: [Identification of interaction and interaction domains between neuroglobin and Na(+), K(+)-ATPase beta2 subunit].
abstract: 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.
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Wen-Lin
authors.initials: WL
authors.lastname: Xu
authors.forename: Chun-Li
authors.initials: CL
authors.lastname: Wang
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Liao
authors.forename: Yong-Liang
authors.initials: YL
authors.lastname: Zhang
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Yu
authors.forename: Fan-Wei
authors.initials: FW
authors.lastname: Meng
authors.forename: Xin-Xing
authors.initials: XX
authors.lastname: Wang
authors.forename: Fan-Wei
authors.initials: FW
authors.lastname: Meng
authors.forename: Zhao-Yun
authors.initials: ZY
authors.lastname: Yin
authors.forename: Ling-Jia
authors.initials: LJ
authors.lastname: Qian
authors.forename: Cheng-Gang
authors.initials: CG
authors.lastname: Zhang
mesh_terms: 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
chemical_list: D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D005914:Globins; D005982:Glutathione Transferase; D000254:Sodium-Potassium-Exchanging ATPase