Identification of red blood cell membrane defects in a patient with hereditary spherocytosis using next‑generation sequencing technology and matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry

Şener,Leyla  Türker; Aktan,Melih; Albeniz,Gürcan; Şener,Aziz; Üstek,Duran; Albeniz,Işıl

doi:10.3892/mmr.2019.10036

Identification of red blood cell membrane defects in a patient with hereditary spherocytosis using next‑generation sequencing technology and matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry

Authors:
- Leyla Türker Şener
- Melih Aktan
- Gürcan Albeniz
- Aziz Şener
- Duran Üstek
- Işıl Albeniz
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Affiliations: Department of Biophysics, Istanbul Faculty of Medicine, Istanbul University, 34093 Istanbul, Turkey, Department of Hematology, Istanbul Faculty of Medicine, Istanbul University, 34093 Istanbul, Turkey, Department of General Surgery, Cerrahpaşa Faculty of Medicine, Istanbul University Cerrahpaşa, 34096 Istanbul, Turkey, Department of General Surgery, Kanuni Sultan Suleyman Training and Research Hospital, 34303 Istanbul, Turkey, Department of Medical Genetics and REMER, Medipol University, 34810 Istanbul, Turkey
Published online on: March 14, 2019 https://doi.org/10.3892/mmr.2019.10036
Pages: 3912-3922

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Abstract

Hereditary spherocytosis (HS) is characterized by the morphological transformation of erythrocytes into a spherical shape due to a hereditary defect in cell membrane proteins (ghosts) associated with disruption of erythrocyte skeletal structures. Contrary to the literature, pores were detected in the erythrocytes of a patient with HS. The aim of the present study was to determine the affected proteins and genes that were responsible for the pores. Ghost isolation was performed to determine the proteins responsible for the pores observed on the erythrocytes of the patient. Erythrocyte membrane proteins were visualized using SDS‑PAGE. Exome and matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry (MALDI TOF MS) analyses were used to identify the genes and proteins responsible for the observed defect. Quantitative protein assessments were performed using MALDI TOF MS. A difference was detected in the components of the erythrocyte membrane proteins. Band 3 and protein 4.2, which serve a particular role in membrane structure, decreased 4.573 and 4.106 fold, respectively. Through proteomic analyses, a non‑synonymous exonic mutation region was identified in the Golgi membrane protein 1 (GOLM1) gene (Chr9 rs142242230). Sorting Intolerant From Tolerant and Polymorphism Phenotyping Scores, Likelihood Ratio Tests and MutationTaster revealed that the mutation was deleterious. The pores observed in the morphology of the erythrocytes may have developed due to the decrease in these proteins, which reside in the erythrocyte membrane structure. Furthermore, genetic profiling of the patient with HS and her family was conducted in the present study. Next‑generation sequencing was used, and the genetic source of HS was identified as a GOLM1 gene mutation. The assessment of specific molecular defects is often not performed as the majority of mutations are unique to a family. However, molecular analyses should be performed in severe cases where prenatal diagnosis is required, or for unique HS phenotypes to aid scientific investigation.

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