ABCMdb

PMID: 19383836

Toyoda Y, Sakurai A, Mitani Y, Nakashima M, Yoshiura K, Nakagawa H, Sakai Y, Ota I, Lezhava A, Hayashizaki Y, Niikawa N, Ishikawa T
Earwax, osmidrosis, and breast cancer: why does one SNP (538G>A) in the human ABC transporter ABCC11 gene determine earwax type?
FASEB J. 2009 Jun;23(6):2001-13. Epub 2009 Apr 21., [PubMed]

Sentences

No. Mutations Sentence Comment

1 ABCC11 p.Gly180Arg ABCC11 p.Gly180Arg Yu Toyoda,* Aki Sakurai,*,† Yasumasa Mitani,†,‡ Masahiro Nakashima,§ Koh-ichiro Yoshiura,ʈ Hiroshi Nakagawa,* Yasuo Sakai,¶ Ikuko Ota,†,# Alexander Lezhava,† Yoshihide Hayashizaki,† Norio Niikawa,ʈ, ** and Toshihisa Ishikawa*,†,1 *Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan; † Omics Science Center (OSC), RIKEN Yokohama Institute, Yokohama, Japan; ‡ K.K. DNAFORM, Yokohama, Japan; § Tissue and Histopathology Section, Atomic Bomb Disease Institute, and ʈ Department of Human Genetics, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; ¶ Department of Plastic and Reconstructive Surgery, Fujita Health University, Toyoake, Japan; # Department of Gastroenterological Surgery, Yokohama City University Graduate School of Medicine, Yokohama, Japan; and **Research Institute of Personalized Health Sciences, Health Sciences University of Hokkaido, Hokkaido, Japan ABSTRACT One single-nucleotide polymorphism (SNP), 538G>A (Gly180Arg), in the ABCC11 gene determines the type of earwax. Login to comment 16 ABCC11 p.Gly180Arg Although the biochemical and physiological pathways that regulate the apocrine secretory process are not clearly known, our recent finding that the nonsynonymous SNP 538GϾA (rs17822931; Gly180Arg) in the ABCC11 gene determines the type of earwax has shed light on the novel function of this ABC transporter in apocrine glands (6). Login to comment 23 ABCC11 p.Gly180Arg We have recently reported that one single-nucleotide polymorphism (SNP), 538GϾA (Gly180Arg), in the ABCC11 gene determines the type of earwax (6). Login to comment 31 ABCC11 p.Gly180Arg First, to understand the molecular mechanism determining the earwax type, we investigated the effect of the SNP 538GϾA (Gly180Arg) on the protein expression and intracellular localization of ABCC11. Login to comment 33 ABCC11 p.Gly180Arg In this article, we provide evidence that the SNP 538GϾA (Gly180Arg) variant of human ABCC11 lacking N-linked glycosylation is recognized as a misfolded protein in the endoplasmic reticulum (ER) and readily undergoes proteasomal degradation. Login to comment 72 ABCC11 p.Gly180Arg Generation of ABCC11 variant forms The human ABCC11 WT or G180R cDNA was inserted into the pcDNA3.1/Hygro(-) vector (Invitrogen) between the restriction enzyme sites of XhoI/SalI and HindIII, respectively. Login to comment 73 ABCC11 p.Gly180Arg ABCC11 p.Asn838Gln ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln ABCC11 p.Asn844Gln The resulting expression construct [ABCC11 WT-pcDNA3.1/ Hygro(-)] was used as the template for site-directed mutagenesis to obtain ABCC11 variants, i.e., N838Q, N844Q, and N838Q/N844Q. Login to comment 74 ABCC11 p.Asn838Gln ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln ABCC11 p.Asn844Gln The resulting expression construct [ABCC11 WT-pcDNA3.1/ Hygro(afa;)] was used as the template for site-directed mutagenesis to obtain ABCC11 variants, i.e., N838Q, N844Q, and N838Q/N844Q. Login to comment 75 ABCC11 p.Asn838Gln ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln To generate the N838Q/N844Q variant, ABCC11 N838Q- pcDNA3.1/Hygro(-) was used as the template. Login to comment 76 ABCC11 p.Asn838Gln ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln To generate the N838Q/N844Q variant, ABCC11 N838Q- pcDNA3.1/Hygro(afa;) was used as the template. Login to comment 77 ABCC11 p.Gly180Arg ABCC11 p.Gly180His ABCC11 p.Gly180Lys To substitute Gly180 to Arg, Lys, His, Asp, Glu, AL, or Pro in the ABCC11 WT protein, the codon (GGG) encoding the Gly residue in TM1 was changed by site-directed mutagenesis, as described above. Login to comment 78 ABCC11 p.Gly180Arg ABCC11 p.Gly180His ABCC11 p.Gly180Lys To substitute Gly180 to Arg, Lys, His, Asp, Glu, AL, or Pro in the ABCC11 WT protein, the codon (GGG) encoding the Gly residue in TM1 was changed by site-directed mutagenesis, as described above. Login to comment 116 ABCC11 p.Gly180Arg It is likely that the nonsynonymous SNP 538GϾA (Gly180Arg) greatly affects the cellular localization of the ABCC11 protein in secretory cells. Login to comment 117 ABCC11 p.Gly180Arg It is likely that the nonsynonymous SNP 538Gb0e;A (Gly180Arg) greatly affects the cellular localization of the ABCC11 protein in secretory cells. Login to comment 126 ABCC11 p.Gly180Arg B) Human ABCC11 WT or G180R cDNA was inserted into pcDNA3.1/Hygro(-) vector between restriction enzyme sites of XhoI/SalI and HindIII, respectively. Login to comment 127 ABCC11 p.Gly180Arg B) Human ABCC11 WT or G180R cDNA was inserted into pcDNA3.1/Hygro(afa;) vector between restriction enzyme sites of XhoI/SalI and HindIII, respectively. Login to comment 161 ABCC11 p.Gly180Arg Since Gly180 or Arg180 residue resides in TM1, this amino acid alteration (Gly180Arg) does not affect immunoreactivity. Login to comment 162 ABCC11 p.Gly180Arg Since Gly180 or Arg180 residue resides in TM1, this amino acid alteration (Gly180Arg) does not affect immunoreactivity. Login to comment 165 ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln Substitution of both Asn838 and Asn844 to Gln residues completely diminished N-linked glycosylation of the ABCC11 WT (Fig. 5B), demonstrating that these two Asn residues are N-linked glycosylation sites in the ABCC11 WT protein. Login to comment 166 ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln Substitution of both Asn838 and Asn844 to Gln residues completely diminished N-linked glycosylation of the ABCC11 WT (Fig. 5B), demonstrating that these two Asn residues are N-linked glycosylation sites in the ABCC11 WT protein. Login to comment 167 ABCC11 p.Gly180Arg ABCC11 p.Gly180His ABCC11 p.Gly180Lys To examine our hypothesis, we substituted Gly180 to Arg, Lys, His, Asp, Glu, AL, and Pro in the ABCC11 WT protein. Login to comment 168 ABCC11 p.Gly180Arg ABCC11 p.Gly180His ABCC11 p.Gly180Lys To examine our hypothesis, we substituted Gly180 to Arg, Lys, His, Asp, Glu, AL, and Pro in the ABCC11 WT protein. Login to comment 172 ABCC11 p.Gly180Arg Both the rare mutation and G180R provide the dry type of earwax (6). Login to comment 173 ABCC11 p.Gly180Arg ABCC11 p.Gly180Arg Among the variants tested, only G180R and ⌬27 diminished N-linked glycosylation of ABCC11 (Fig. 6B). Login to comment 174 ABCC11 p.Gly180Arg Among the variants tested, only G180R and èc;27 diminished N-linked glycosylation of ABCC11 (Fig. 6B). Login to comment 176 ABCC11 p.Gly180Arg Clinical genotyping of the SNP 538G>A (Gly180Arg) in the ABCC11 gene by the SmartAmp method We tried to create a clinical method to genotype the SNP 538GϾA in the heman ABCC11 gene. Login to comment 177 ABCC11 p.Gly180Arg Clinical genotyping of the SNP 538G>A (Gly180Arg) in the ABCC11 gene by the SmartAmp method We tried to create a clinical method to genotype the SNP 538Gb0e;A in the heman ABCC11 gene. Login to comment 178 ABCC11 p.Gly180Arg Effect of SNP 538GϾA (Gly180Arg) on expression of human ABCC11 protein in Flp-In-293 cells. Login to comment 179 ABCC11 p.Gly180Arg Effect of SNP 538Gb0e;A (Gly180Arg) on expression of human ABCC11 protein in Flp-In-293 cells. Login to comment 193 ABCC11 p.Gly180Arg To determine the SNP 538GϾA (Gly180Arg) in the ABCC11 gene, we prepared one set of primers designated TP, FP, BP, OP, and CP (Fig. 7B). Login to comment 194 ABCC11 p.Gly180Arg To determine the SNP 538Gb0e;A (Gly180Arg) in the ABCC11 gene, we prepared one set of primers designated TP, FP, BP, OP, and CP (Fig. 7B). Login to comment 198 ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln B) Effect of substitution of Asn838 and Asn844 to Gln on N-linked glycosylation status of ABCC11. Login to comment 199 ABCC11 p.Asn838Gln ABCC11 p.Asn844Gln B) Effect of substitution of Asn838 and Asn844 to Gln on N-linked glycosylation status of ABCC11. Login to comment