ABCMdb

PMID: 10615958

Tanizawa Y, Matsuda K, Matsuo M, Ohta Y, Ochi N, Adachi M, Koga M, Mizuno S, Kajita M, Tanaka Y, Tachibana K, Inoue H, Furukawa S, Amachi T, Ueda K, Oka Y
Genetic analysis of Japanese patients with persistent hyperinsulinemic hypoglycemia of infancy: nucleotide-binding fold-2 mutation impairs cooperative binding of adenine nucleotides to sulfonylurea receptor 1.
Diabetes. 2000 Jan;49(1):114-20., [PubMed]

Sentences

No. Mutations Sentence Comment

2 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys In the SUR1 gene, we identified one frameshift (I446fsdelT) and two missense (R1420C, R1436Q) mutations. Login to comment 4 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Siblings homozygous for the R1420C mutation had a mild form, whereas two patients heterozygous for the I446fsdelT and R1436Q mutations, respectively, exhibited a severe form of PHHI. Login to comment 62 ABCC8 p.Arg1420Cys The I446fsdelT and R1420C mutations created an NlaIII restriction site (CATG). Login to comment 63 ABCC8 p.Arg1436Gln The R1436Q mutation created a BsrI restriction site (CCAGTG). Login to comment 64 ABCC8 p.Arg1436Gln After PCR amplification of exon9 (I446fsdelT)or exon 35 (R1420Cand R1436Q), the products were digested with the appropriate restriction enzymes and analyzed by electrophoresis on 2% agarose gel. Login to comment 66 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Site-directed mutagenesis was performed using mutagenesis primers (for I446fsdelT: 5 -acg ctc cgt tca tgc ctc tcc atc atc c-3 ; for R1420C: 5 -gcc agt aca gat cat gtg ggc gtg atc ctc c-3 ; and for R1436Q: 5 -ttc agc ggc acc atc caa ttc aacctg gac cc-3 ) and a GeneEditor in vitro Site-DirectedMuta- genesisSystem (Promega, Madison, WI). Login to comment 67 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Site-directed mutagenesis was performed using mutagenesis primers (for I446fsdelT: 5 -acg ctc cgt tca tgc ctc tcc atc atc c-3 ; for R1420C: 5 -gcc agt aca gat cat gtg ggc gtg atc ctc c-3 ; and for R1436Q: 5 -ttc agc ggc acc atc caa ttc aacctg gac cc-3 ) and a GeneEditor in vitro Site-DirectedMuta- genesisSystem (Promega, Madison, WI). Login to comment 80 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys We identified three potential disease-causing mutations (I446fsdelT, R1420C, and R1436Q). Login to comment 81 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys We identified three potential disease-causing mutations (I446fsdelT, R1420C, and R1436Q). Login to comment 89 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Two other mutations (R1420C and R1436Q) were missense mutations, both in NBF-2, a functionally important domain of SUR1. Login to comment 90 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Two other mutations (R1420C and R1436Q) were missense mutations, both in NBF-2, a functionally important domain of SUR1. Login to comment 92 ABCC8 p.Arg1420Cys Two patients (patients 2 and 3), siblings from a consanguineous family, were homozygous for the R1420C mutation. Login to comment 93 ABCC8 p.Arg1420Cys Two patients (patients 2 and 3), siblings from a consanguineous family, were homozygous for the R1420C mutation. Login to comment 94 ABCC8 p.Arg1436Gln R1436Q is a novel missense mutation. Login to comment 95 ABCC8 p.Arg1436Gln R1436Q is a novel missense mutation. Login to comment 110 ABCC8 p.Arg1420Cys A sibling pair (patients 2 and 3), who were homozygous for the R1420C mutation, had a milder form of PHHI. Login to comment 111 ABCC8 p.Arg1420Cys A sibling pair (patients 2 and 3), who were homozygous for the R1420C mutation, had a milder form of PHHI. Login to comment 119 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Therefore, we introduced I446fsdelT, R1420C, and R1436Q mutations by site-directed mutagenesis to the corresponding positions of mouse SUR1 cDNA, and mutant SUR1 and mouse Kir6.2 were transiently coexpressed in COS-7 cells to reconstitute the mutant KATP channel. Login to comment 120 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys Therefore, we introduced I446fsdelT, R1420C, and R1436Q mutations by site-directed mutagenesis to the corresponding positions of mouse SUR1 cDNA, and mutant SUR1 and mouse Kir6.2 were transiently coexpressed in COS-7 cells to reconstitute the mutant KATP channel. Login to comment 123 ABCC8 p.Arg1436Gln Because the R1436Q mutation is a missense mutation, the protein expression level of SUR1-1436Q was assessed by Western blot analysis. Login to comment 124 ABCC8 p.Arg1436Gln Because the R1436Q mutation is a missense mutation, the protein expression level of SUR1-1436Q was assessed by Western blot analysis. Login to comment 127 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys ABCC8 p.Arg1420Cys Activation ofthe channel by diazoxide also appeared to be impaired in the KATP(SUR1-1420C) mutant, TABLE 3 Profiles of patients with SUR1 gene mutations Onset Birth Treatment/ Patient Mutation (day) weight (g) outcome 1 I446fsdelT 0 5,014 Partial pancreatectomy 2 R1420C 0 5,254 Remission 3 R1420C 0 5,080 Remission 4 R1436Q 0 3,410 Partial pancreatectomy FIG. 1. Login to comment 128 ABCC8 p.Arg1436Gln ABCC8 p.Arg1420Cys ABCC8 p.Arg1420Cys Activation ofthe channel by diazoxide also appeared to be impaired in the KATP(SUR1-1420C) mutant, TABLE 3 Profiles of patients with SUR1 gene mutations Onset Birth Treatment/ Patient Mutation (day) weight (g) outcome 1 I446fsdelT 0 5,014 Partial pancreatectomy 2 R1420C 0 5,254 Remission 3 R1420C 0 5,080 Remission 4 R1436Q 0 3,410 Partial pancreatectomy FIG. 1. Login to comment 130 ABCC8 p.Arg1420Cys Means of measurements made in triplicate for the representative experiment are plotted. , SUR1 (wild-type); , SUR1-R1420C; , SUR1-446delT; , LacZ. Login to comment 131 ABCC8 p.Arg1420Cys Means of measurements made in triplicate for the representative experiment are plotted. , SUR1 (wild-type); , SUR1-R1420C; , SUR1-446delT; , LacZ. Login to comment 140 ABCC8 p.Arg1420Cys Effect of R1420C mutation on cooperative binding of adenine nucleotides in SUR1. There is a cooperative regulation of ATP and ADP binding to SUR1, and this cooperativity may be involved in MgATP and MgADP regulation of the KATP channel (18,19). Login to comment 143 ABCC8 p.Arg1420Cys Effects of the R1420C mutation in NBF-2 on cooperative binding of ATP and ADP were assessed. Login to comment 160 ABCC8 p.Arg1436Gln This may also be the case for patient 4, who was heterozygous for a germline R1436Q mutation. Login to comment 196 ABCC8 p.Arg1420Cys This may also be the case for the R1420C mutation, although we did not test this possibility directly. Login to comment 199 ABCC8 p.Arg1420Cys ABCC8 p.Arg1436Cys In our 86 Rb+ efflux studies, impairment of KATP channel function was modest with the R1420C mutation, while no channel activities were observed for the other two mutations, R1436C and I446fsdelT. Login to comment 200 ABCC8 p.Arg1420Cys ABCC8 p.Arg1436Cys This parallels clinical disease severities of these patients: patients with the R1420C mutation achieved seemingly spontaneous remission after a few months of medical therapy, whereas patients who had R1436C or I446fsdelT mutations required partial pancreatectomy. Login to comment