Substitution Of Arginine For Glycine Research Articles

Brothers with novel compound heterozygous mutations in COL27A1 causing dental and genital abnormalities

COL27A1 encodes a collagen type XXVII alpha 1 chain. It is the product of this gene that provides the structural support of connective tissue and is reported to be the causative gene of Steel syndrome (OMIM #615155). The primary symptoms of patients with this defect are consistent with systemic bone disease; however, recent reports note findings of intellectual disability and hearing loss. In this study, we identified novel COL27A1 compound heterozygous variants in two brothers with rhizomelia and congenital hip dislocation as well as dental and genital abnormalities that have not yet been reported in Steel syndrome. This variant, of maternal origin, caused an amino acid substitution of arginine for glycine, c.2026G>C or p.G676R, in the collagen helix domain, which is assumed to damage the structure of the helix. The paternally transmitted variant, c.2367G>A, is located at the 3′ end of exon 12, and cDNA analysis revealed a splicing alteration. These novel, compound heterozygous COL27A1 variants might indicate an association of the gene with tooth and genital abnormalities.

Abstract 5856: Association between FGFR4 Gly388Arg polymorphism rs351855 or the IDH1 rs11554137 and Japanese gastric cancer

Abstract Introduction: In recent years, certain single-nucleotide polymorphisms (SNPs) have been reported to be associated with increased risk of cancer and prognosis. A Fibroblast growth factor receptor 4 (FGFR4) Gly388Arg polymorphism (rs351855) which caused the substitution of arginine for glycine in the transmembrane domain of the receptor, was reported to increase cancer risk or not. The meta-analysis results showed that FGFR4 Gly388Arg polymorphism (rs351855) might be a cancer risk factor for majority Asians, especially in breast cancer and prostate cancer. There is also a report that FGFR4 Gly388Arg polymorphism (rs351855) may be a prognostic biomarker for lung squamous-cell carcinoma patients with lymph node metastasis and uterine cervical cancer. But few studies have been conducted in gastric cancer. And the Isocitrate Dehydrogenase 1 (IDH1) mutations in patients with acute myelogenous leukemia (AML) who have a normal karyotype are associated with a poor prognosis. IDH1 rs11554137:C>T, which is located in codon 105 in the same exon as the IDH1 codon 132 arginine (IDH1R132) mutation, has been described in patients with AML and has been identified as an adverse prognostic factor. It has also been reported that IDH1 rs11554137:C>T has a negative effect on glioblastoma patients in terms of both progression free survival and overall survival (OS). We investigated the association of the gastric cancer with two SNPs. Methods: This study included 30 cases and the tumor samples obtained from gastric cancer patients by surgery between October 2010 and December 2018. We evaluated all 30 samples by a panel testing (OncoGuide NCC Oncopanel system). And we searched for the expression of FGFR4 Gly388Arg polymorphism (rs351855) and the IDH1 rs11554137. Univariate analysis and Kaplan-Meier analysis was performed based on the OS. Comparison between two groups was evaluated using chi-squared test and Fisher's exact test. The stage evaluation of the cancer adopted Japanese Gastric Cancer Association the 15th Edition. Results: Median age is 70 (34-86) years, gender is Male/Female=18/12 cases, histology is Intestinal/diffuse/Scirrhous=8/13/9 cases. There were 21 cases of FGFR4 Gly388Arg polymorphism (rs351855) expression and 2 cases of the IDH1 rs11554137 expression in 30 cases of gastric cancer. The expression of FGFR4 Gly388Arg polymorphism (rs351855) and the IDH1 rs11554137 was compared with various clinic-pathologic factors, however, no significant correlation was found between clinic-pathologic factors and FGFR4 Gly388Arg polymorphism (rs351855) and the IDH1 rs11554137 expression. Conclusion: Our results suggest that neither FGFR4 Gly388Arg polymorphism (rs351855) nor IDH1 rs11554137 is not associated with the progression of gastric cancer. Citation Format: Tomohiro Sera, Masakazu Yashiro, Yurie Yamamoto, Yukako Kushitani, Atsushi Sugimoto, Syuhei Kushiyama, Kenji Kuroda, Shingo Togano, Tomohisa Okuno, Mami Yoshii, Tatsuro Tamura, Takahiro Toyokawa, Hiroaki Tanaka, Kazuya Muguruma, Masaichi Ohira. Association between FGFR4 Gly388Arg polymorphism rs351855 or the IDH1 rs11554137 and Japanese gastric cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5856.

Association between FGFR4 Gly388Arg polymorphism (rs351855) and cancer risk: A meta analysis including 10,584 subjects

A FGFR4 Gly388Arg polymorphism (rs351855) which caused the substitution of arginine for glycine in the transmembrane domain of the receptor, was reported to increase cancer risk or not. According to this reason, a meta analysis was made, including a total of 4767 cases and 5817 controls. The results showed that FGFR4 Gly388Arg polymorphism (rs351855) might be an cancer risk factor for majority Asians, especially in breast cancer and prostate cancer. But it was necessary to complete several large samples studies of various type cancers from different ethnic populations.

Whole‑exome sequencing identifies a novel mutation (R367G) in SCN5A to be associated with familial cardiac conduction disease.

Cardiac conduction disease is a primary cause of sudden cardiac death. Sodium voltage‑gated channel‑α subunit5 (SCN5A) mutations have been reported to underlie a variety of inherited arrhythmias. Numerous disease‑causing mutations of SCN5A have been identified in patients with ≥10 different conditions, including type3 long‑QT syndrome and Brugada syndrome. The present study investigated a family with a history of arrhythmia, with the proband having a history of arrhythmia and syncope. Whole‑exome sequencing was applied in order to detect the disease‑causing mutation in this family, and Sanger sequencing was used to confirm the co‑segregation among the family members. A missense mutation (c.1099C>G/p.R367G) of SCN5A was identified in the family and was observed to be co‑segregated in all affected members of the family. The missense mutation results in a substitution of glycine for arginine, which may affect sodium transmembrane transport. The present study provides an accurate genetic test which may be used in individuals who exhibit no clinical symptoms.

Single amino acid substitution in homogentisate 1,2-dioxygenase is responsible for pigmentation in a subset of Burkholderia cepacia complex isolates.

The Burkholderia cepacia complex (Bcc) is a group of Gram-negative bacilli that are ubiquitous in the environment and have emerged over the past 30 years as opportunistic pathogens in immunocompromised populations, specifically individuals with cystic fibrosis (CF) and chronic granulomatous disease. This complex of at least 18 distinct species is phenotypically and genetically diverse. One phenotype observed in a subset of Burkholderia cenocepacia (a prominent Bcc pathogen) isolates is the ability to produce a melanin-like pigment. Melanins have antioxidant properties and have been shown to act as virulence factors allowing pathogens to resist killing by the host immune system. The melanin-like pigment expressed by B. cenocepacia is produced through tyrosine catabolism, specifically through the autoxidation and polymerization of homogentisate. Burkholderia cenocepacia J2315 is a CF clinical isolate that displays a pigmented phenotype when grown under normal laboratory conditions. We examined the amino acid sequences of critical enzymes in the melanin synthesis pathway in pigmented and non-pigmented Bcc isolates, and found that an amino acid substitution of glycine for arginine at amino acid 378 in homogentisate 1,2-dioxygenase correlated with pigment production; we identify this as one mechanism for expression of pigment in Bcc isolates.

G127R: A novel SOD1 mutation associated with rapidly evolving ALS and severe pain syndrome

We describe a patient with apparently sporadic amyotrophic lateral sclerosis (SALS) with a novel g > c point mutation at position 382 in the SOD1 gene, leading to a substitution of glycine for arginine in amino acid position 127 (G127R). The disease presented with flaccid leg paresis, and progressed rapidly with generalized paresis resulting in respiratory failure after seven months. In addition to a predominating lower motor neuron syndrome, the phenotype was characterized by a severe lower back and leg pain syndrome which was treated successfully with spinal anaesthesia.

Dwarf 88, a novel putative esterase gene affecting architecture of rice plant

Rice architecture is an important agronomic trait that affects grain yield. We characterized a tillering dwarf mutant d88 derived from Oryza sativa ssp. japonica cultivar Lansheng treated with EMS. The mutant had excessive shorter tillers and smaller panicles and seeds compared to the wild-type. A reduction in number and size of parenchyma cells around stem marrow cavity as well as a delay in the elongation of parenchyma cells caused slender tillers and dwarfism in the d88 mutant. The D88 gene was isolated via map-based cloning and identified to encode a putative esterase. The gene was expressed in most rice organs, with especially high levels in the vascular tissues. The mutant carried a nucleotide substitution in the first exon of the gene that led to the substitution of arginine for glycine, which presumably disrupted the functionally conserved N-myristoylation domain of the protein. The function of the gene was confirmed by complementation test and antisense analysis. D88, thus, represents a new category of genes that regulates cell growth and organ development and consequently plant architecture. The potential relationship between the tiller formation associated genes and D88 is discussed and future identification of the substrate for D88 may lead to the characterization of new pathways regulating plant development.

Influence of β2-Adrenergic Receptor Genotype on Airway Function During Exercise in Healthy Adults

In humans, beta(2)-adrenergic receptors (beta(2)ARs) influence airway tone. There are known functional polymorphisms of the beta(2)AR, such as substitution of glycine for arginine at codon 16. We sought to determine if this variation in genotype differentially influences airway function during exercise. Healthy subjects without asthma who were either homozygous for Arg16 (n = 16; mean age, 29 +/- 2 years [+/- SD]; mean maximum oxygen uptake [Vo(2)], 32 +/- 2 mL/kg/min) or the Gly16 allele (n = 26; mean age, 30 +/- 1 years; mean maximum Vo(2), 33 +/- 1 mL/kg/min) participated in the study. Baseline testing included spirometry and maximal symptom-limited exercise. On a separate day, an arterial cannula was placed to measure catecholamine levels. Subjects then performed exercise at two work levels (40% and 75% of peak work) for 9 min each and performed spirometry at 3-min intervals for assessment of airway function. There were no statistically significant differences between groups in maximum Vo(2) or baseline spirometry (p > 0.05). With both light and heavy exercise, the groups had similar increases in the forced expiratory flow at 50% of vital capacity (FEF(50)). FEF(50) increased by 14 +/- 4% and 15 +/- 3% in arginine and glycine groups, respectively, by end exercise (p > 0.05). During recovery (5 min and 10 min after), the Gly16 homozygotes demonstrated persistent bronchodilation (10 min after FEF(50) = + 7 +/- 2% over pre-exercise) while the Arg16 subjects had a rapid return to baseline (10 min after FEF(50) = - 3 +/- 3%, p = 0.007 between groups). No differences were observed in the catecholamine responses between genotypes, although the increase in epinephrine in the arginine group tended to be higher (p = 0.07). These data suggest that the Arg16Gly polymorphism of the beta(2)AR does not influence airway function during short-duration low- and high-intensity exercise. However, during recovery, the Arg16 genotype is associated with a reduced bronchodilation, possibly due to increased catecholamine desensitization.

Targeted introduction of a diphtheria toxin resistant mutation into the chromosomal EF-2 locus of Pichia pastoris and expression of immunotoxin in the EF-2 mutants

In an attempt to increase the production of a diphtheria toxin (DT) based immunotoxin by Pichia pastoris, we have created DT-resistant mutants that contain a substitution of arginine for glycine at position 701 in elongation factor 2 (EF-2). To achieve this, we first cloned and characterized the EF-2 gene ( PEF1), and then made a construct pBLURA-Δ 5 ′ mutEF-2 that efficiently introduces specific mutations into the chromosomal EF-2 gene in P. pastoris by in vivo homologous recombination. pBLURA-Δ 5 ′ mutEF-2 contains a selection marker URA3 and a 5 ′ truncated form of the P. pastoris PEF1 that had been modified in vitro to carry the nucleotide mutations for the Gly 701 to Arg transition. Unlike the non-mutated strains, the EF-2 mutants are resistant to high-level intracellular expression of DT A chain that can catalyze the ADP-ribosylation. When used to express the secreted bivalent anti-T cell immunotoxin, A-dmDT390-bisFv(G4S), the EF-2 mutant strains showed increased viability compared to the non-mutated strains. However, they did not show an advantage over the non-mutated expressing strain in the production of the immunotoxin. Western blotting analysis revealed that although the EF-2 mutants did not increase the accumulation of intact A-dmDT390-bisFv(G4S) in the culture medium, they generated larger amounts of degraded products found in both the medium and cell pellets compared to the non-mutant expressing clone. In addition, double copy expression resulted in greater amounts of intact immunotoxin being retained within cellular compartments as well as degraded products. Based on these findings, we suggest that the secretory capacity may be rate limiting for divalent immunotoxin production in P. pastoris.

Welcome to the machine: a cardiologist's introduction to protein folding and degradation.

The price paid by the cardiovascular system for maintaining circulation is being a target of chronic forces that contribute to wear and tear. Cardiovascular tissues are in constant peril of damage from these stresses, and are dependent on highly conserved cellular machines for protection. The cellular environment is adapted to detect, repair and, if necessary, dispose of damaged proteins, in part because they are toxic to the cell. The molecular chaperone and ubiquitin-proteasome systems are the machinery that repair and degrade damaged proteins, yet little attention has been given to these systems in cardiovascular pathophysiology. Recent advances have brought protein folding and degradation closer to the forefront of cardiovascular biology. This review discusses these advances and presents new models for consideration of cardiovascular pathophysiology and therapeutics as problems of protein homeostasis. In most cases, all the information necessary for a protein to fold is contained in its primary amino acid sequence. The environment within a cell is particularly unfavorable for protein folding, however, because of constraints that occur during protein synthesis and the likelihood of intermolecular interactions that impede folding in a crowded cellular environment. An evolutionarily ancient system exists to prevent damaged or newly synthesized peptides from aggregating and to provide a microenvironment that facilitates their proper folding to a thermodynamically favorable active conformation. This system comprises the molecular chaperones, which are present in every cellular compartment to buffer and repair damaged proteins (the Table provides a partial list of mammalian chaperones). Mechanisms for removing proteins are equally critical to cellular function, and the machinery of protein degradation is similarly archaic. Although several pathways exist for protein destruction in eukaryotes, the ubiquitin-proteasome system is responsible for the majority of protein degradation and is the most tightly regulated pathway. View this table: Examples of Mammalian Chaperones and Their Functions The chaperone and ubiquitin-proteasome …

Skeletal development of achondroplasia: analysis of genotyped patients.

Achondroplasia is a skeletal dysplasia caused by substitution of arginine for glycine at codon 380 (G380R) mutation of the fibroblast growth factor receptor 3. To date, the developmental course of the phenotype (short stature and skeletal characteristics) has not been clarified in the genotyped population. The relationship between age and clinical data (height, arm span and measurements of skeletal radiographs) were statistically analyzed from 27 achondroplasia patients with the G380R genotype. The height standard deviation score had positive correlation and decreased with age, while span-to-height ratio did not. Among measurements of skeletal radiographs, the pelvic index, which represents the squared pelvis deformity, were correlated and increased with age. However, interpedicular distance of the first and fourth lumbar vertebrae (L1:L4) ratio as an index for the caudally narrowed pattern of the lumbar spinal canal and fibula-to-tibia ratio for the disproportionally long fibulae were not correlated and did not increase with age. In making a clinical diagnosis of achondroplasia in early infancy, it should be noted that short stature and squared pelvis deformity are not prominent.

Analyses of the transglutaminase 1 gene mutation and ultrastructural characteristics in a Japanese patient with lamellar ichthyosis

We described a Japanese female with lamellar ichthyosis whose transglutaminase 1 gene (TGM1 gene) was mutated. DNA sequence analysis revealed that the patient had a homozygous mutation, i.e. a point mutation from G to A at nucleotide 1494 resulting in the substitution of glycine for arginine at codon 143. Her mother was heterozygous for this mutation. In situ transglutaminase assay in the patient's skin showed loss of enzyme activity. Ultrastructural examination revealed incomplete formation of cornified cell envelopes and electron-dense materials adjacent to plasma membranes. These results suggest that defective transglutaminase activity caused by homozygous TGM1 gene mutation (G143R) results in disruption of cornified envelope assembly and the clinical phenotype of lamellar ichthyosis.

Mutation in the gene encoding the fibroblast growth factor receptor-3 in Korean children with achondroplasia.

Achondroplasia (ACH) is the most common form of osteochondrodysplasia, and is mostly associated with a point mutation in the gene on the transmembrane domain of fibroblast growth factor receptor-3 (FGFR-3) on chromosome 4p. We investigated the mutations in the gene encoding FGFR-3 in 15 Korean children with ACH, using polymerase chain reaction (PCR) coupled with direct sequencing. In this study, all children with ACH showed the same mutation as those reported in France, USA and Japan; a G-->A transition at position 1138 of the coding sequence, resulting in the substitution of arginine for glycine at position 380 of the mature protein. This consistent point mutation of Korean children with ACH indicates there is no significant racial difference in the pathogenesis of ACH, compared with data from Caucasian and Japanese children with ACH.

Modification of a Synthetic Antimicrobial Peptide (ESF1) for Improved Inhibitory Activity

Rational modification of an existing cationic α-helical antimicrobial peptide (ESF1) for improved activity by increasing amphipathicity was undertaken. ESF1 and two variants (GR7 and SA3) were synthesized and tested for activity range, minimum inhibitory concentration, and hemolytic activity. Biological activity was related to structure as determined by circular dichroism. The substitution of arginine for glycine in position seven was found to increase antimicrobial activity without effecting hemolysis. Increased activity was related to stronger α-helix formation in buffer. Increased β-sheet formation in micellar SDS was observed and speculated to be due to a stronger ability of the variants to form multimolecule complexes, a feature consistent with existing models of cationic peptide activity.

A mutation in the lipoprotein lipase gene is the molecular basis of chylomicronemia in a colony of domestic cats.

Members of a domestic cat colony with chylomicronemia share many phenotypic features with human lipoprotein lipase (LPL) deficiency. Biochemical analysis reveals that these cats do have defective LPL catalytic activity and have a clinical phenotype very similar to human LPL deficiency. To determine the molecular basis underlying this biochemical phenotype, we have cloned the normal and affected cat LPL cDNAs and shown that the affected cat has a nucleotide change resulting in a substitution of arginine for glycine at residue 412 in exon 8. In vitro mutagenesis and expression studies, in addition to segregation analysis, have shown that this DNA change is the cause of LPL deficiency in this cat colony. Reduced body mass, growth rates, and increased stillbirth rates are observed in cats homozygous for this mutation. These findings show that this LPL deficient cat can serve as an animal model of human LPL deficiency and will be useful for in vivo investigation of the relationship between triglyceride rich lipoproteins and atherogenic risk and for the assessment of new approaches for treatment of LPL deficiency, including gene therapy.

Substitution of arginine for glycine at position 154 of the α1 chain of type I collagen in a variant of osteogenesis imperfecta: Comparison to previous cases with the same mutation

A substitution of arginine for glycine at amino acid position 154 of the alpha 1(I) collagen chain was found in a father and his three children. The phenotype of the patients includes manifestations of types I and III/IV osteogenesis imperfecta, but appears to be milder than that of the previously described two unrelated patients that had the identical mutation in the alpha 1(I) collagen chain. The variability in the phenotype raises the possibility of epistatic loci or environmental effects on expression of the disorder.

Substitution of arginine for glycine at position 154 of the α1 chain of type I collagen in a variant of osteogenesis imperfecta: Comparison to previous cases with the same mutation

A substitution of arginine for glycine at amino acid position 154 of the alpha 1(I) collagen chain was found in a father and his three children. The phenotype of the patients includes manifestations of types I and III/IV osteogenesis imperfecta, but appears to be milder than that of the previously described two unrelated patients that had the identical mutation in the alpha 1(I) collagen chain. The variability in the phenotype raises the possibility of epistatic loci or environmental effects on expression of the disorder.

Identification of Mutations in the Gene Encoding the Fibroblast Growth Factor Receptor 3 in Japanese Patients with Achondroplasia

Abstract We examined mutations in the gene encoding the fibroblast growth factor receptor 3 (FGFR3) in nine Japanese patients diagnosed as typical achondroplasia (ACH) and two patients with hypochondroplasia (HCH) using polymerase chain reaction (PCR) coupled with direct sequencing. In our present cases, equivalent to previously reported cases in France and the U. S. A., all mutations affected with ACH were the same nucleotide change, a G→A transition at position 1,138, resulting in the substitution of arginine for glycine at position 380 of the mature protein. Therefore, the gene mutation of ACH indicates no significant differences between the Caucasian and the Japanese. On the other hand, PCR products from genomic DNA of HCH did not show the same mutation.

Fulminant reactivation of hepatitis B due to envelope protein mutant that escaped detection by monoclonal HBsAg ELISA

The detection of a fatal case of reactivation of hepatitis B, in a previously vaccinated Indonesian patient after withdrawal of chemotherapy for lymphoma, was delayed because HBsAg was negative in a widely used monoclonal- antibody-based ELISA. The serum was later found to be strongly reactive for HBsAg by the polyclonal radioimmunoassay and for HBV DNA. PCR sequencing revelated a substitution of arginine for glycine at position 145 of HBsAg in the major neutralising epitope cluster, the a determinant, as well as a 2–aminoacid insertion of asparagine and threonine between positions 122 and 123, immediately upstream of this determinant.

Three New Exon 10 Glucose-6-Phosphate Dehydrogenase Mutations

ABSTRACT Three previously undescribed mutations of the glucose-6-phosphate dehydrogenase (G6PD) gene have been documented in patients with hereditary non-spherocytic hemolytic anemia (HNSHA). In none of the cases have we been able to obtain a sufficient volume of blood to characterize the residual enzyme biochemically. "G6PD Calvo Mackenna" was due to an A_G transition in cDNA nucleotide 1138 creating an Aat II site and resulting in a substitution of valine for isoleucine at amino acid 380. "G6PD Riley" was due to a T_C transition at cDNA nucleotide 1139 also changing the 380 isoleucine, in this case to a threonine. "G6PD Wisconsin" was due to an C_G transversion in cDNA nucleotide 1177, destroying a Aci I site and resulting in a substitution of glycine for arginine at amino acid 393. All of these mutations were in exon 10, where mutations that cause HNSHA appear to be clustered. We present a list of the 83 mutations of G6PD that have been documented to the end of April, 1995.