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A novel missense mutation in the ALPL gene causes dysfunction of the protein

  • Authors:
    • Bin Chen
    • Lili Li
    • Weitong Ren
    • Long Yi
    • Yaping Wang
    • Fuhua Yan

  • View Affiliations

  • Published online on: May 31, 2017     https://doi.org/10.3892/mmr.2017.6668
  • Pages: 710-718

  • Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypophosphatasia (HP) is a rare genetic disease caused by mutation in the alkaline phosphatase, liver/bone/kidney (ALPL) gene with highly variable clinical manifestations. Efforts have been made to collect cases with novel mutations and to examine how a missense mutation affects ALPL protein function, which remains difficult to predict. The present study investigated the underlying mechanism of ALPL dysfunction in a patient diagnosed with HP. Bidirectional sequencing of the ALPL gene was conducted in a 5‑year‑old Chinese girl preliminary diagnosed with childhood HP. Sorting Intolerant from Tolerant (SIFT) and Polymorphism Phenotyping v2 (PolyPhen‑2) tools were used to forecast the impact of the mutation on protein function. Site‑directed mutagenesis was performed and transfected into cells to verify the role of the specific mutation. Furthermore, the mechanism of the impact was investigated via all‑atom molecular dynamics (MD) simulation. The patient demonstrated a compound heterozygote with two missense mutations in the ALPL gene, p.Trp29Arg and p.Ile395Val. Trp29 and Ile395 were determined to be ‘tolerable’ by SIFT, whereas they were ‘possibly damaging’ by PolyPhen‑2 in terms of conservation. Additionally, HEK293 cells were transfected with plasmids expressing wild type and/or mutated ALPL. Only 4.1% of ALP activity remained when Trp29 was substituted by Arg, whereas 19.1, 33.7, 50.1 and 7.6% ALP activity remained in cells expressing p.Ile395Val, wild type+p.Trp29Arg, wild type+p.Ile395Val and p.Trp29Arg+p.Ile395Val substitutions, respectively. All‑atom MD simulation demonstrated that the N‑terminal helix of mutated ALPL, where Trp29 is located, separated from the main body of the protein after 30 nsec, and moved freely. These results demonstrated that p.Trp29Arg, as a novel missense mutation in the ALPL gene, reduced the enzymatic activity of ALPL. This effect may be associated with an uncontrolled N‑terminal helix. These results provide novel information about the genetic basis of HP, and may facilitate the development of future therapies.
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July-2017
Volume 16 Issue 1

Print ISSN: 1791-2997
Online ISSN:1791-3004

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  • CiteScore: 7.6
  • CiteScore Rank: Q1: #82/347 Biochemistry, Genetics and Molecular Biology
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Spandidos Publications style
Chen B, Li L, Ren W, Yi L, Wang Y and Yan F: A novel missense mutation in the ALPL gene causes dysfunction of the protein. Mol Med Rep 16: 710-718, 2017.
APA
Chen, B., Li, L., Ren, W., Yi, L., Wang, Y., & Yan, F. (2017). A novel missense mutation in the ALPL gene causes dysfunction of the protein. Molecular Medicine Reports, 16, 710-718. https://doi.org/10.3892/mmr.2017.6668
MLA
Chen, B., Li, L., Ren, W., Yi, L., Wang, Y., Yan, F."A novel missense mutation in the ALPL gene causes dysfunction of the protein". Molecular Medicine Reports 16.1 (2017): 710-718.
Chicago
Chen, B., Li, L., Ren, W., Yi, L., Wang, Y., Yan, F."A novel missense mutation in the ALPL gene causes dysfunction of the protein". Molecular Medicine Reports 16, no. 1 (2017): 710-718. https://doi.org/10.3892/mmr.2017.6668