PMID- 36979346
OWN - NLM
STAT- MEDLINE
DCOM- 20230330
LR  - 20231213
IS  - 2218-273X (Electronic)
IS  - 2218-273X (Linking)
VI  - 13
IP  - 3
DP  - 2023 Feb 22
TI  - KI04 an Aminoglycosides-Derived Molecule Acts as an Inhibitor of Human Connexin46 
      Hemichannels Expressed in HeLa Cells.
LID - 10.3390/biom13030411 [doi]
LID - 411
AB  - BACKGROUND: Connexins (Cxs) are proteins that help cells to communicate with the 
      extracellular media and with the cytoplasm of neighboring cells. Despite their 
      importance in several human physiological and pathological conditions, their 
      pharmacology is very poor. In the last decade, some molecules derived from 
      aminoglycosides have been developed as inhibitors of Cxs hemichannels. However, 
      these studies have been performed in E. coli, which is a very simple model. 
      Therefore, our main goal is to test whether these molecules have similar effects 
      in mammalian cells. METHODS: We transfected HeLa cells with the human Cx46tGFP 
      and characterized the effect of a kanamycin-derived molecule (KI04) on Cx46 
      hemichannel activity by time-lapse recordings, changes in phosphorylation by 
      Western blot, localization by epifluorescence, and possible binding sites by 
      molecular dynamics (MD). RESULTS: We observed that kanamycin and KI04 were the 
      most potent inhibitors of Cx46 hemichannels among several aminoglycosides, 
      presenting an IC(50) close to 10 muM. The inhibitory effect was not associated 
      with changes in Cx46 electrophoretic mobility or its intracellular localization. 
      Interestingly, 5 mM DTT did not reverse KI04 inhibition, but the KI04 effect 
      completely disappeared after washing out KI04 from the recording media. MD 
      analysis revealed two putative binding sites of KI04 in the Cx46 hemichannel. 
      RESULTS: These results demonstrate that KI04 could be used as a Cx46 inhibitor 
      and could help to develop future selective Cx46 inhibitors.
FAU - Chang, Cheng-Wei T
AU  - Chang CT
AUID- ORCID: 0000-0002-8978-4520
AD  - Department of Chemistry and Biochemistry, Utah State University, Logan, UT 
      84322-0300, USA.
FAU - Poudyal, Naveena
AU  - Poudyal N
AD  - Department of Chemistry and Biochemistry, Utah State University, Logan, UT 
      84322-0300, USA.
FAU - Verdugo, Daniel A
AU  - Verdugo DA
AUID- ORCID: 0000-0003-0256-454X
AD  - Laboratorio de Neurobiologia, Facultad de Medicina y Facultad de Ciencias de la 
      Vida, Instituto de Ciencias Biomedicas, Universidad Andres Bello, Santiago 
      7780272, Chile.
FAU - Pena, Francisca
AU  - Pena F
AD  - Laboratorio de Neurobiologia, Facultad de Medicina y Facultad de Ciencias de la 
      Vida, Instituto de Ciencias Biomedicas, Universidad Andres Bello, Santiago 
      7780272, Chile.
FAU - Stehberg, Jimmy
AU  - Stehberg J
AD  - Laboratorio de Neurobiologia, Facultad de Medicina y Facultad de Ciencias de la 
      Vida, Instituto de Ciencias Biomedicas, Universidad Andres Bello, Santiago 
      7780272, Chile.
FAU - Retamal, Mauricio A
AU  - Retamal MA
AUID- ORCID: 0000-0003-2562-6686
AD  - Center for Membrane Protein Research, Department of Cell Physiology and Molecular 
      Biophysics, Texas Tech University Health Sciences Center, Lubbock, TX 79430-6551, 
      USA.
AD  - Programa de Comunicacion Celular en Cancer, Facultad de Medicina Clinica Alemana, 
      Universidad del Desarrollo, Santiago 7610496, Chile.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20230222
PL  - Switzerland
TA  - Biomolecules
JT  - Biomolecules
JID - 101596414
RN  - 0 (GJA3 protein, human)
RN  - 0 (Aminoglycosides)
RN  - 0 (Connexins)
RN  - 0 (Anti-Bacterial Agents)
RN  - 59-01-8 (Kanamycin)
SB  - IM
MH  - Animals
MH  - Humans
MH  - HeLa Cells
MH  - *Aminoglycosides
MH  - *Escherichia coli/metabolism
MH  - Connexins/metabolism
MH  - Anti-Bacterial Agents
MH  - Kanamycin/pharmacology
MH  - Mammals/metabolism
PMC - PMC10046693
OTO - NOTNLM
OT  - aminoglycosides
OT  - cancer
OT  - connexins
OT  - hemichannels
OT  - inhibitors
OT  - lens
COIS- The authors declare no conflict of interest.
EDAT- 2023/03/30 06:00
MHDA- 2023/03/30 06:11
PMCR- 2023/02/22
CRDT- 2023/03/29 01:19
PHST- 2023/01/21 00:00 [received]
PHST- 2023/02/12 00:00 [revised]
PHST- 2023/02/16 00:00 [accepted]
PHST- 2023/03/30 06:11 [medline]
PHST- 2023/03/29 01:19 [entrez]
PHST- 2023/03/30 06:00 [pubmed]
PHST- 2023/02/22 00:00 [pmc-release]
AID - biom13030411 [pii]
AID - biomolecules-13-00411 [pii]
AID - 10.3390/biom13030411 [doi]
PST - epublish
SO  - Biomolecules. 2023 Feb 22;13(3):411. doi: 10.3390/biom13030411.