PMID- 28498321 OWN - NLM STAT- MEDLINE DCOM- 20180222 LR - 20181113 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 18 IP - 5 DP - 2017 May 12 TI - BDNF Binds Its Pro-Peptide with High Affinity and the Common Val66Met Polymorphism Attenuates the Interaction. LID - 10.3390/ijms18051042 [doi] LID - 1042 AB - Most growth factors are initially synthesized as precursors then cleaved into bioactive mature domains and pro-domains, but the biological roles of pro-domains are poorly understood. In the present study, we investigated the pro-domain (or pro-peptide) of brain-derived neurotrophic factor (BDNF), which promotes neuronal survival, differentiation and synaptic plasticity. The BDNF pro-peptide is a post-processing product of the precursor BDNF. Using surface plasmon resonance and biochemical experiments, we first demonstrated that the BDNF pro-peptide binds to mature BDNF with high affinity, but not other neurotrophins. This interaction was more enhanced at acidic pH than at neutral pH, suggesting that the binding is significant in intracellular compartments such as trafficking vesicles rather than the extracellular space. The common Val66Met BDNF polymorphism results in a valine instead of a methionine in the pro-domain, which affects human brain functions and the activity-dependent secretion of BDNF. We investigated the influence of this variation on the interaction between BDNF and the pro-peptide. Interestingly, the Val66Met polymorphism stabilized the heterodimeric complex of BDNF and its pro-peptide. Furthermore, compared with the Val-containing pro-peptide, the complex with the Met-type pro-peptide was more stable at both acidic and neutral pH, suggesting that the Val66Met BDNF polymorphism forms a more stable complex. A computational modeling provided an interpretation to the role of the Val66Met mutation in the interaction of BDNF and its pro-peptide. Lastly, we performed electrophysiological experiments, which indicated that the BDNF pro-peptide, when pre-incubated with BDNF, attenuated the ability of BDNF to inhibit hippocampal long-term depression (LTD), suggesting a possibility that the BDNF pro-peptide may interact directly with BDNF and thereby inhibit its availability. It was previously reported that the BDNF pro-domain exerts a chaperone-like function and assists the folding of the BDNF protein. However, our results suggest a new role for the BDNF pro-domain (or pro-peptide) following proteolytic cleave of precursor BDNF, and provide insight into the Val66Met polymorphism. FAU - Uegaki, Koichi AU - Uegaki K AD - Biomedical Research Institute (BMD), National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorioka, Ikeda, Osaka 563-8577, Japan. k-uegaki@aist.go.jp. AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. k-uegaki@aist.go.jp. FAU - Kumanogoh, Haruko AU - Kumanogoh H AD - Biomedical Research Institute (BMD), National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorioka, Ikeda, Osaka 563-8577, Japan. h.kumanogoh@gmail.com. AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. h.kumanogoh@gmail.com. FAU - Mizui, Toshiyuki AU - Mizui T AD - Biomedical Research Institute (BMD), National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorioka, Ikeda, Osaka 563-8577, Japan. t-mizui@aist.go.jp. AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. t-mizui@aist.go.jp. FAU - Hirokawa, Takatsugu AU - Hirokawa T AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. t-hirokawa@aist.go.jp. AD - Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Tokyo 135-0064, Japan. t-hirokawa@aist.go.jp. AD - Division of Biomedical Science, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki 305-8575, Japan. t-hirokawa@aist.go.jp. FAU - Ishikawa, Yasuyuki AU - Ishikawa Y AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. yishikaw@maebashi-it.ac.jp. AD - Department of Systems Life Engineering, Maebashi Institute of Technology 460-1, Kamisadori, Maebashi 370-0816, Japan. yishikaw@maebashi-it.ac.jp. FAU - Kojima, Masami AU - Kojima M AD - Biomedical Research Institute (BMD), National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorioka, Ikeda, Osaka 563-8577, Japan. m-kojima@aist.go.jp. AD - Core Research for Evolutional Science and Technology (CREST), Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan. m-kojima@aist.go.jp. AD - Graduate School of Frontier Bioscience, Osaka University, Suita 565-0871, Japan. m-kojima@aist.go.jp. LA - eng PT - Journal Article DEP - 20170512 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Brain-Derived Neurotrophic Factor) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/chemistry/genetics/*metabolism/pharmacology MH - Hippocampus/drug effects/physiology MH - Humans MH - Long-Term Synaptic Depression/drug effects MH - Male MH - Mice MH - Mice, Inbred C57BL MH - *Mutation, Missense MH - *Polymorphism, Single Nucleotide MH - Protein Binding MH - Protein Multimerization MH - Proteolysis PMC - PMC5454954 OTO - NOTNLM OT - BDNF OT - hippocampus OT - long-term depression OT - polymorphism OT - pro-peptide COIS- The authors declare no conflict of interest. EDAT- 2017/05/13 06:00 MHDA- 2018/02/23 06:00 PMCR- 2017/05/01 CRDT- 2017/05/13 06:00 PHST- 2017/04/07 00:00 [received] PHST- 2017/05/06 00:00 [revised] PHST- 2017/05/08 00:00 [accepted] PHST- 2017/05/13 06:00 [entrez] PHST- 2017/05/13 06:00 [pubmed] PHST- 2018/02/23 06:00 [medline] PHST- 2017/05/01 00:00 [pmc-release] AID - ijms18051042 [pii] AID - ijms-18-01042 [pii] AID - 10.3390/ijms18051042 [doi] PST - epublish SO - Int J Mol Sci. 2017 May 12;18(5):1042. doi: 10.3390/ijms18051042.