Chemical conjugation of DeltaF508-CFTR corrector deoxyspergualin to transporter human serum albumin enhances its ability to rescue Cl- channel functions.
The most common mutation of the cystic fibrosis (CF) gene, the deletion of Phe508, encodes a protein (DeltaF508-CFTR) that fails to fold properly, thus mutated DeltaF508-cystic fibrosis transmembrane conductance regulator (CFTR) is recognized and degraded via the ubiquitin-proteasome endoplasmic reticulum-associated degradation pathway. Chemical and pharmacological chaperones and ligand-induced transport open options for designing specific drugs to control protein (mis)folding or transport. A class of compounds that has been proposed as having potential utility in DeltaF508-CFTR is that which targets the molecular chaperone and proteasome systems. In this study, we have selected deoxyspergualin (DSG) as a reference molecule for this class of compounds and for ease of cross-linking to human serum albumin (HSA) as a protein transporter. Chemical cross-linking of DSG to HSA via a disulfide-based cross-linker and its administration to cells carrying DeltaF508-CFTR resulted in a greater enhancement of DeltaF508-CFTR function than when free DSG was used. Function of the selenium-dependent oxidoreductase system was required to allow intracellular activation of HSA-DSG conjugates. The principle that carrier proteins can deliver pharmacological chaperones to cells leading to correction of defective CFTR functions is therefore proven and warrants further investigations.
Norez
Caroline
C
Laboratory of Molecular Pathology, Laboratory of Clinical Chemistry and Hematology, University Hospital of Verona, Verona, Italy.
Pasetto
Matteo
M
Dechecchi
Maria Cristina
MC
Barison
Erika
E
Anselmi
Cristina
C
Tamanini
Anna
A
Quiri
Federica
F
Cattel
Luigi
L
Rizzotti
Paolo
P
Dosio
Franco
F
Cabrini
Giulio
G
Colombatti
Marco
M
D002460:Cell Line; D003432:Cross-Linking Reagents / Q000737:chemistry; D003550:Cystic Fibrosis / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology*; D006146:Guanidines / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology*; D018832:Molecular Chaperones / Q000737:chemistry / Q000494:pharmacology*; D010088:Oxidoreductases / Q000378:metabolism; D017354:Point Mutation; D017510:Protein Folding; D012709:Serum Albumin / Q000737:chemistry / Q000494:pharmacology*
C505032:CFTR protein, human; D003432:Cross-Linking Reagents; D004220:Disulfides; D004337:Drug Carriers; D006146:Guanidines; D007166:Immunosuppressive Agents; D018832:Molecular Chaperones; D012709:Serum Albumin; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D010088:Oxidoreductases; C037258:gusperimus