Sequencing and expression of a full‐length cDNA for human<i>α</i>‐L‐fucosidase

Fukushima, Hisao; Nishimoto, Junji; Okada, Shintaro

doi:10.1007/bf01799583

Cited by 6 publications

(3 citation statements)

References 7 publications

(7 reference statements)

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“…Several bacterial and mammalian α-fucosidases have been biochemically characterized (Fukushima et al 1991;Sulzenbacher et al 2004). Among fungi, a secreted α-fucosidase was purified from the filamentous fungus Fusarium oxysporum, but the encoding gene was not identified (Yamamoto et al 1986).…”

Section: Introductionmentioning

confidence: 99%

α-Fucosidases with different substrate specificities from two species of Fusarium

Paper

Scott-Craig

Cavalier

et al. 2012

Appl Microbiol Biotechnol

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Two fungal-secreted α-fucosidases and their genes were characterized. FoFCO1 was purified from culture filtrates of Fusarium oxysporum strain 0685 grown on L-fucose and its encoding gene identified in the sequenced genome of strain 4287. FoFCO1 was active on p-nitrophenyl-α-fucoside (pNP-Fuc), but did not defucosylate a nonasaccharide (XXFG) fragment of pea xyloglucan. A putative α-fucosidase gene (FgFCO1) from Fusarium graminearum was expressed in Pichia pastoris. FgFCO1 was ~1,800 times less active on pNP-Fuc than FoFCO1, but was able to defucosylate the XXFG nonasaccharide. Although FgFCO1 and FoFCO1 both belong to Glycosyl Hydrolase family 29, they share <25 % overall amino acid identity. Alignment of all available fungal orthologs of FoFCO1 and FgFCO1 indicated that these two proteins belong to two subfamilies of fungal GH29 α-fucosidases. Fungal orthologs of subfamily 1 (to which FoFCO1 belongs) are taxonomically more widely distributed than subfamily 2 (FgFCO1), but neither was universally present in the sequenced fungal genomes. Trichoderma reesei and most species of Aspergillus lack genes for either GH29 subfamily.

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Section: Introductionmentioning

confidence: 99%

α-Fucosidases with different substrate specificities from two species of Fusarium

Paper

Scott-Craig

Cavalier

et al. 2012

Appl Microbiol Biotechnol

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“…A plasmid containing the full-length, wild-type, C~-L-fUcosidase eDNA [pSVL-FUC (wt)] was provided by Hisao Fukushima, Osaka University, Osaka, Japan (Fukushima et al 1990). Site-directed mutagenesis for construction of pSVL-FUC (A86~ was performed according to Deng and Nicholoff (1992).…”

Section: Methodsmentioning

confidence: 99%

A Gln-281 to Arg substitution in ?-l-fucosidase is responsible for a common polymorphism detected by isoelectric focusing

Yang

DiCioccio

1994

Hum Genet

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A common polymorphism of human alpha-L-fucosidase consists of three phenotypes (Fu 1, Fu 2, and Fu 2-1) assigned by isoelectric focusing. The phenotypes are determined by two codominant alleles (Fu1 and Fu2). Isozymes with the Fu 2 phenotype have more basic pI than Fu 1, while Fu 2-1 is a mixture of Fu 2 and Fu 1. Recently, a missense mutation (A860-->G) in the alpha-L-fucosidase gene was described that did not affect alpha-L-fucosidase activity. The mutation causes the substitution of Arg (pKaGuan = 12.5) for Gln-281, which has no ionizable side chain. Isoelectric focusing profiles of extracts of COS-1 cells transfected with wild-type or mutant alpha-L-fucosidase cDNAs had phenotypes of Fu 1 and Fu 2, respectively. Next, 20 human lymphoid cell lines were examined for the occurrence of the A860-->G mutation and expression of the Fu 1, Fu 2, and Fu 2-1 phenotypes. Eight lines with Fu 2 were homozygous for the A860-->G mutation; six lines with Fu 1 were homozygous for the normal nucleotide (A860); and six lines with Fu 2-1 were heterozygous. Thus, the A860-->G mutation is the molecular basis for the protein phenotypes and the Fu1 and Fu2 alleles. The normal nucleotide (A860) is responsible for Fu1 and the A860-->G mutation for Fu2.

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“…The human a-L-fucosidase gene (FUCA1) spans over 23 kb, contains 8 exons, and has been mapped to chromosome 1p34.1-1p36.1 (Fukushima et al 1985(Fukushima et al , 1990Occhiodoro et al 1989;Kretz et al 1992). The 2,053 bp full-length cDNA encodes for a signal peptide of 22 amino acids and a mature protein of 439 amino acids.…”

Section: Introductionmentioning

confidence: 99%

Mutation identification and characterization of a Taiwanese patient with fucosidosis

et al. 2007

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Fucosidosis is a rare lysosomal storage disease caused by a defect of the a-L-fucosidase (FUCA1) gene. Worldwide 26 mutations underlying the disease have been reported. By direct DNA sequencing of exons and flanking introns, homozygous Y126X mutation and Q281R polymorphism were found in a Taiwanese patient with fucosidosis. Upon expressing in COS-7 cells, 97.4% of a-Lfucosidase activity compared with that of the wild-type construct was observed in the cDNA containing Q281R polymorphism. Western blot analysis revealed a 58-kDa precursor and 56-kDa mature forms for cells transfected with wild-type and Q281R enzymes. Using the fluorogenic substrate, the Michaelis constants and maximal velocities of both enzymes were very similar. While no appreciable enzyme activity (0.0%) was observed with Y126X mutation, no apparent decrease in FUCA1 mRNA level was seen with Y126X mutation. The expressed truncated Y126X protein was unstable and largely degraded. The delineation of the molecular defect could serve to complement future prenatal diagnosis for this family when necessary.

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Sequencing and expression of a full‐length cDNA for humanα‐L‐fucosidase

Cited by 6 publications

References 7 publications

α-Fucosidases with different substrate specificities from two species of Fusarium

α-Fucosidases with different substrate specificities from two species of Fusarium

A Gln-281 to Arg substitution in ?-l-fucosidase is responsible for a common polymorphism detected by isoelectric focusing

Mutation identification and characterization of a Taiwanese patient with fucosidosis

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