Sains Malaysiana 53(3)(2024): 681-689

http://doi.org/10.17576/jsm-2024-5303-15

Investigation of Phosphorylated and O-Glycosylated Proteins in Gestational Diabetes Mellitus

(Penelitian Protein Terfosforilasi dan Terglikosilasi O dalam Kehamilan Diabetes Mellitus)

AFSHAN KALEEM*, TEHREEM ANWAR, AQSA IFTIKHAR, ROHEENA ABDULLAH, MEHWISH IQTEDAR & MAHWISH AFTAB

Department of Biotechnology, Lahore College for Women University, Lahore-54000, Pakistan

Received: 17 August 2023/Accepted: 21 February 2024

Abstract

Gestational Diabetes Mellitus (GDM) is a common but temporary type of diabetes that develops among 10-20% of all pregnant women. It is a major cause of several pre- and post-pregnancy diseases in both mother and offspring. The main causative is altered pregnancy hormones that lead to deficient glucose metabolism. Complications can be preeclampsia, caesarean sections and cardiovascular diseases. This study was designed to determine the phosphorylated and glycosylated proteins present in GDM and to investigate the role of their post-translational modifications (PTMs) in the pathogenesis of GDM. The study was performed on 30 blood samples from pregnant diabetic women (diseased), 30 pregnant non-diabetic women (control) and 30 women without pregnancy and gestational diabetes (normal). Protein extraction was done from blood plasma, and phosphorylated and glycosylated proteins were determined by Enzyme-Linked Immuno-Sorbent Assay (ELISA) by use of specific antibodies. This study showed that proteins found in GDM are highly phosphorylated and O-glycosylated (p<0.01). Furthermore, a bioinformatic investigation was done, which showed that blood coagulation proteins such as the Fibrinogen alpha chain and lipid metabolism-regulating proteins apolipoprotein E, L1 and C-III showed a high potential for phosphorylation, which suggests that phosphorylation can be used as a therapeutic marker in GDM.

Keywords: Gestational diabetes mellitus; glycosylation; phosphorylation; protein modification

Abstrak

Kehamilan Diabetes Mellitus (GDM) adalah jenis diabetes biasa tetapi sementara yang berlaku dalam kalangan 10-20% daripada semua wanita hamil. Ia adalah punca utama kepada beberapa penyakit sebelum dan selepas kehamilan pada ibu dan anak. Penyebab utama adalah perubahan hormon kehamilan yang membawa kepada kekurangan metabolisme glukosa. Komplikasi boleh menjadi preeklampsia, pembedahan caesarean dan penyakit kardiovaskular. Kajian ini direka bentuk untuk menentukan protein terfosforilasi dan terglikosilasi yang hadir dalam GDM dan untuk mengkaji peranan pengubahsuaian pasca translasi (PTM) mereka dalam patogenesis GDM. Kajian dilakukan terhadap 30 sampel darah daripada wanita hamil yang menghidap diabetes (berpenyakit), 30 wanita hamil bukan kencing manis (kawalan) dan 30 wanita tidak hamil dan tiada kencing manis (normal). Pengekstrakan protein dilakukan daripada plasma darah dan protein terfosforilasi dan glikosilasi ditentukan oleh Asai Imunosorben Berkait Enzim (ELISA) dengan menggunakan antibodi khusus. Kajian ini menunjukkan bahawa protein yang terdapat dalam GDM adalah sangat terfosforilasi dan O-glikosilasi (p<0.01). Tambahan pula, penyelidikan bioinformatik telah dilakukan yang menunjukkan bahawa protein pembekuan darah seperti rantai alfa Fibrinogen dan protein yang mengawal metabolisme lipid apolipoprotein E, L1 dan C-III menunjukkan potensi tinggi untuk fosforilasi yang mencadangkan bahawa fosforilasi boleh digunakan sebagai penanda terapeutik dalam GDM.

Kata kunci: Kehamilan Diabetes Mellitus; pengubahsuaian protein; glikosilasi; fosforilasi

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*Corresponding author; email: afshan.kaleem@lcwu.edu.pk