PMID- 35970844 OWN - NLM STAT- MEDLINE DCOM- 20220817 LR - 20220912 IS - 2049-3169 (Electronic) IS - 1674-2818 (Print) IS - 1674-2818 (Linking) VI - 14 IP - 1 DP - 2022 Aug 15 TI - GPCR/endocytosis/ERK signaling/S2R is involved in the regulation of the internalization, mitochondria-targeting and -activating properties of human salivary histatin 1. PG - 42 LID - 10.1038/s41368-022-00181-5 [doi] LID - 42 AB - Human salivary histatin 1 (Hst1) exhibits a series of cell-activating properties, such as promoting cell spreading, migration, and metabolic activity. We recently have shown that fluorescently labeled Hst1 (F-Hst1) targets and activates mitochondria, presenting an important molecular mechanism. However, its regulating signaling pathways remain to be elucidated. We investigated the influence of specific inhibitors of G protein-coupled receptors (GPCR), endocytosis pathways, extracellular signal-regulated kinases 1/2 (ERK1/2) signaling, p38 signaling, mitochondrial respiration and Na+/K+-ATPase activity on the uptake, mitochondria-targeting and -activating properties of F-Hst1. We performed a siRNA knockdown (KD) to assess the effect of Sigma-2 receptor (S2R) /Transmembrane Protein 97 (TMEM97)-a recently identified target protein of Hst1. We also adopted live cell imaging to monitor the whole intracellular trafficking process of F-Hst1. Our results showed that the inhibition of cellular respiration hindered the internalization of F-Hst1. The inhibitors of GPCR, ERK1/2, phagocytosis, and clathrin-mediated endocytosis (CME) as well as siRNA KD of S2R/TMEM97 significantly reduced the uptake, which was accompanied by the nullification of the promoting effect of F-Hst1 on cell metabolic activity. Only the inhibitor of CME and KD of S2R/TMEM97 significantly compromised the mitochondria-targeting of Hst1. We further showed the intracellular trafficking and targeting process of F-Hst1, in which early endosome plays an important role. Overall, phagocytosis, CME, GPCR, ERK signaling, and S2R/TMEM97 are involved in the internalization of Hst1, while only CME and S2R/TMEM97 are critical for its subcellular targeting. The inhibition of either internalization or mitochondria-targeting of Hst1 could significantly compromise its mitochondria-activating property. CI - (c) 2022. The Author(s). FAU - Ma, Dandan AU - Ma D AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. AD - School of Stomatology, Zhejiang Chinese Medical University, Hangzhou, China. AD - Laboratory for Myology, Department of Human Movement Sciences, Faculty of Behavioral and Movement Sciences, Amsterdam Movement Sciences, Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Sun, Wei AU - Sun W AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Fu, Cuicui AU - Fu C AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Nazmi, Kamran AU - Nazmi K AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Veerman, Enno C I AU - Veerman ECI AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Jaspers, Richard T AU - Jaspers RT AD - Laboratory for Myology, Department of Human Movement Sciences, Faculty of Behavioral and Movement Sciences, Amsterdam Movement Sciences, Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Bolscher, Jan G M AU - Bolscher JGM AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Bikker, Floris J AU - Bikker FJ AD - Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Wu, Gang AU - Wu G AUID- ORCID: 0000-0001-8941-2500 AD - Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam（ACTA）, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, Netherlands. g.wu@acta.nl. AD - Department of Oral and Maxillofacial Surgery/Pathology, Amsterdam UMC and Academic Center for Dentistry Amsterdam （ACTA）, Vrije Universiteit Amsterdam, Amsterdam Movement Science, Amsterdam, Netherlands. g.wu@acta.nl. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220815 PL - India TA - Int J Oral Sci JT - International journal of oral science JID - 101504351 RN - 0 (Histatins) RN - 0 (Membrane Proteins) RN - 0 (RNA, Small Interfering) RN - 0 (Receptors, G-Protein-Coupled) RN - 0 (Receptors, sigma) RN - 0 (TMEM97 protein, human) RN - 0 (sigma-2 receptor) RN - 101056-53-5 (HTN1 protein, human) SB - IM MH - *Endocytosis/physiology MH - *Histatins/pharmacology MH - Humans MH - Membrane Proteins MH - Mitochondria/metabolism MH - RNA, Small Interfering/pharmacology MH - Receptors, G-Protein-Coupled/metabolism MH - Receptors, sigma PMC - PMC9378733 COIS- The authors declare no competing interests. EDAT- 2022/08/16 06:00 MHDA- 2022/08/18 06:00 PMCR- 2022/08/15 CRDT- 2022/08/15 23:15 PHST- 2021/12/27 00:00 [received] PHST- 2022/05/10 00:00 [accepted] PHST- 2022/04/14 00:00 [revised] PHST- 2022/08/15 23:15 [entrez] PHST- 2022/08/16 06:00 [pubmed] PHST- 2022/08/18 06:00 [medline] PHST- 2022/08/15 00:00 [pmc-release] AID - 10.1038/s41368-022-00181-5 [pii] AID - 181 [pii] AID - 10.1038/s41368-022-00181-5 [doi] PST - epublish SO - Int J Oral Sci. 2022 Aug 15;14(1):42. doi: 10.1038/s41368-022-00181-5.