Valosin containing protein (VCP) interacts with macrolide antibiotics without mediating their anti-inflammatory activities.
In addition to antibacterial activity, some macrolide antibiotics, such as azithromycin and clarithromycin, also exhibit anti-inflammatory properties in vitro and in vivo, although the targets and mechanism(s) of action remain unknown. The aim of the present study was to identify protein targets of azithromycin and clarithromycin which could potentially explain their anti-inflammatory effects. Using chemical proteomics approach, based on compound-immobilized affinity chromatography, valosin containing protein (VCP) was identified as a potential target of the macrolides. Validation studies confirmed the interaction of macrolides and VCP and gave some structural characteristics of this interaction. Cell based assays however, including the use of gene silencing and the study of VCP specific cellular functions in J774.A1 (murine macrophage) and IB3-1 (human cystic fibrotic epithelial) cell lines, failed to confirm an association between the binding of the macrolides to VCP and anti-inflammatory effects. These findings suggest the absence of an abundant high affinity protein target and the potential involvement of other biological molecules in the anti-inflammatory activity of macrolides.
GlaxoSmithKline Research Centre Zagreb, Prilaz baruna Filipovića 29, 10000 Zagreb, Croatia. krunoslav.nujic@glpg.com
Krunoslav
K
Nujić
Marjorie
M
Smith
Michael
M
Lee
Daniela
D
Belamarić
Linda
L
Tomašković
Sulejman
S
Alihodžić
Ivica
I
Malnar
Denis
D
Polančec
Klaus
K
Schneider
Vesna
V
Eraković Haber
D000251:Adenosine Triphosphatases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D000893:Anti-Inflammatory Agents / Q000378:metabolism / Q000494:pharmacology; D017963:Azithromycin / Q000378:metabolism* / Q000494:pharmacology*; D018797:Cell Cycle Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D017291:Clarithromycin / Q000378:metabolism* / Q000494:pharmacology*; D015195:Drug Design; D020868:Gene Silencing; D006801:Humans; D051379:Mice; D011485:Protein Binding; D040901:Proteomics; D015203:Reproducibility of Results; D000074405:Valosin Containing Protein
D000900:Anti-Bacterial Agents; D000893:Anti-Inflammatory Agents; D018797:Cell Cycle Proteins; D017963:Azithromycin; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; C000614179:Vcp protein, mouse; D017291:Clarithromycin