title: [Establishment of an ErbB3- and ErbB4-specific ligand screening system, and a screen of neuregulin-1 mutants].
abstract: Neuregulin-1 (NRG1), now in a phase II clinical trial, has beneficial effects on heart failure patients through the activation of ErbB2/ErbB4 receptor pair. To decrease the side effect of NRG1 on activating ErbB3, a mutation screen was carried out to get NRG1 mutants, which have more specific binding to ErbB2/ErbB4 receptor pair. Two CHO stable cell lines were constructed, which express ErbB2/ErbB3 or ErbB2/ErbB4 receptor pair, respectively. The ErbB2/ErbB4 cell line showed similar characteristics in ligand-binding activity and the activation of downstream signaling molecules, such as the AKT and PI3K to the primary neonatal rat ventricular myocytes (NRVM), which endogenously expresses ErbB2/ErbB4. Both cell lines have good dose-response. Thirty-one NRG1 mutants were successfully expressed in Escherichia coli and purified. Their ability to stimulate the downstream signaling was measured by detecting AKT phosphorylation. Some mutants showed more specific activation activity in ErbB2/ErbB4 cells. Further study on five of these mutants demonstrated that the change of the activation activity is associated with that of their binding activities to ErbB2/ErbB4 and ErbB2/ErbB3. Four of the candidates are more specific ligands for ErbB2/ErbB4 activation, and thus may serve as more potent drug candidates for heart failure.
authors.affiliation: Newworld Institute of Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. qiying99@netease.com
authors.forename: Ying
authors.initials: Y
authors.lastname: Qi
authors.forename: Dong-Zhi
authors.initials: DZ
authors.lastname: Wei
authors.forename: Xi-Fu
authors.initials: XF
authors.lastname: Liu
authors.forename: Ming-Dong
authors.initials: MD
authors.lastname: Zhou
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D066246:ErbB Receptors / Q000378:metabolism*; D006333:Heart Failure / Q000188:drug therapy; D008024:Ligands; D032342:Muscle Cells / Q000378:metabolism; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D009154:Mutation; D020890:Neuregulin-1 / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D015202:Protein Engineering / Q000379:methods*; D051381:Rats; D020893:Receptor, ErbB-3 / Q000378:metabolism*; D066247:Receptor, ErbB-4; D013379:Substrate Specificity; D014162:Transfection
chemical_list: D008024:Ligands; D050505:Mutant Proteins; D020890:Neuregulin-1; D066246:ErbB Receptors; C587691:Erbb4 protein, rat; D020893:Receptor, ErbB-3; D066247:Receptor, ErbB-4