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Effect of ‘attenuated’ mutations in mucopolysaccharidosis IVA on molecular phenotypes of N-acetylgalactosamine-6-sulfate sulfatase

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Summary

Mucopolysaccharidosis IVA is an autosomal recessive disease caused by the deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Mutation screening of the GALNS gene was performed for seven MPS IVA patients with attenuated phenotypes from three unrelated families. Four of 5 missense mutations identified in this study (p.F167V, p.R253W, p.R380S, p.P484S) and two reported (p.F97V, p.N204K), associated with attenuated phenotypes, were characterized using in vitro stable expression experiments, enzyme kinetic study, protein processing and structural analysis. The stably expressed mutant enzymes defining the attenuated phenotype exhibited a considerable residual activity (1.2–36.7% of the wild-type GALNS activity) except for p.R380S. Enzyme kinetic studies showed that p.F97V, p.F167V and p.N204K have lower affinity to the substrate compared with other mutants. The p.F97V enzyme was the most thermolabile at 55°C. Immunoblot analyses indicated a rapid degradation and/or an insufficiency in processing in the mutant proteins. Tertiary structure analysis revealed that although there was a tendency for ‘attenuated’ mutant residues to be located on the surface of GALNS, they have a different effect on the protein including modification of the hydrophobic core and salt-bridge formation and different potential energy. This study demonstrates that ‘attenuated’ mutant enzymes are heterogeneous in molecular phenotypes, including biochemical properties and tertiary structure.

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Authors and Affiliations

  1. Department of Pediatrics, Saint Louis University, Pediatric Research Institute, 3662 Park Ave., St. Louis, MO, 63110-2586, USA

    A. M. Montaño & S. Tomatsu

  2. Department of Pediatrics, Gifu University, Gifu, Japan

    A. M. Montaño, K. Sukegawa, Z. Kato, K. O. Orii, T. Orii & N. Kondo

  3. Servizio di Genetica Medica, Istituto Scientifico Ospedale ‘San Raffaele’, Milan, Italy

    R. Carrozzo

  4. Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Naples, Italy

    P. Di Natale

  5. Department of Clinical Genetics, The Juliane Marie Center, Copenhagen University Hospital, Copenhagen, Denmark

    E. Christensen

Authors
  1. A. M. Montaño
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  2. K. Sukegawa
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  3. Z. Kato
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  4. R. Carrozzo
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  5. P. Di Natale
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  6. E. Christensen
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  7. K. O. Orii
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  8. T. Orii
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  9. N. Kondo
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  10. S. Tomatsu
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Corresponding author

Correspondence to S. Tomatsu.

Additional information

Communicating editor: Ed Wraith

Competing interests: None declared

References to electronic databases: MPS IVA: OMIM #253000. GALNS: EC 3.1.6.4.

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Montaño, A.M., Sukegawa, K., Kato, Z. et al. Effect of ‘attenuated’ mutations in mucopolysaccharidosis IVA on molecular phenotypes of N-acetylgalactosamine-6-sulfate sulfatase. J Inherit Metab Dis 30, 758–767 (2007). https://doi.org/10.1007/s10545-007-0702-z

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  • DOI: https://doi.org/10.1007/s10545-007-0702-z

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