ABCMdb

PMID: 7684377

Chang XB, Tabcharani JA, Hou YX, Jensen TJ, Kartner N, Alon N, Hanrahan JW, Riordan JR
Protein kinase A (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites.
J Biol Chem. 1993 May 25;268(15):11304-11., [PubMed]

Sentences

No. Mutations Sentence Comment

7 ABCC7 p.Ser660Ala ABCC7 p.Ser737Ala ABCC7 p.Ser795Ala A previous investigationconcluded thatactivationby PKA is crit- icallydependent on phosphorylation at four of the nine predicted PKA sites in the R domain (S660A, S737A, S795A,SS13A), becausea "Quad"mutant lacking these sites couldnotbeactivated. Login to comment 37 ABCC7 p.Ser660Ala ABCC7 p.Ser737Glu ABCC7 p.Ser795Glu ABCC7 p.Ser813Glu ABCC7 p.Ser660Glu ABCC7 p.Ser737Ala ABCC7 p.Ser795Ala ABCC7 p.Ser813Ala ABCC7 p.Ser686Ala ABCC7 p.Ser768Ala ABCC7 p.Ser712Ala ABCC7 p.Ser700Ala ABCC7 p.Ser422Ala ABCC7 p.Thr788Ala The following mutations were introduced into CFTR, S422A (TCT to GCT), S660A (TCA to GCA), S686A (TCT to GCT), S700A (TCT toGCT), S712A (TCC to GCC), S737A (TCC to GCC), S768A (TCT toGCT), T788A (ACAto GCA),S795A (TCA to GCA), S813A (TCA to GCA), S660E (TCA to GAA),S737E (TCC to GAG), S795E (TCA to GAA), and S813E (TCA to GAA). Login to comment 39 ABCC7 p.Ser660Ala ABCC7 p.Ser737Ala ABCC7 p.Ser795Ala ABCC7 p.Ser813Ala The counterparts of CFTR cDNA in pUCF2.5 were replaced by PCR- mutated versions by interchange of the following fragments, S660A, DraIIIIEcoRI fragment, S737A, EcoRIIHpaIfragment, S795A and S813A,StyIIStyI fragment(Fig. 1A). Login to comment 40 ABCC7 p.Ser660Ala ABCC7 p.Ser737Ala A mutant containing S660/737/ 795/813A (pUCF2.5/4SA) was assembled by replacing the counterparts of a plasmid containing S795/813A with the DraIII/EcoRI fragment (S660A) and EcoRI/HpaI fragment (S737A). Login to comment 44 ABCC7 p.Ser700Ala S700A (mutation at 2230 bp destroys the EcoRI site) and S712Awere introduced into pUCF2.5/4SA by replacing the DraIIIIHpaI fragment (Fig. lA)containing S660/700/712/737A (the construct was named pUCF2.5/6SA). Login to comment 45 ABCC7 p.Ser768Ala ABCC7 p.Thr788Ala The DraIII/DraIIIfragment in pNUT-CFTR was replaced by the counterpart from pUCF2.5/6SA to generate 6SA.S686A,S768A, and T788Awere introduced into pUCF2.5/6SA by replacing the counterpart of DraIII/HpaI fragment (containing S660/686/700/712/737/768A) andStyIIStyI fragment (containing T788A/S795/813A). Login to comment 47 ABCC7 p.Ser422Ala S422A was introduced into the XbaIIBamHIfragment (Fig. lA)and inserted A into pUCF2.5/9SA. Login to comment 69 ABCC7 p.Ser660Ala For example, S660A means that serine residue 660 was changed to alanine. Login to comment 137 ABCC7 p.Gly551Asp Hence we introduced a mutation at an essential amino acid residue in NBFl known to cause severe disease in patients, i.e. G551D (19). Login to comment 139 ABCC7 p.Gly551Asp Furthermore,cAMPstimulation of phosphorylation of the G551D protein was notdiminished (Fig. 5B). Login to comment