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Purification, Characterization and Binding of Metals to Zinc Alpha 2-Glycoprotein Zag by Employing Various Techniques

Thesis Info

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External Link

Author

Zain Ullah

Program

PhD

Institute

Gomal University

City

Dera Ismail Khan

Province

KPK

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Physical Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10612/1/PhD%20Thesis%20in%20PDF%20formate.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676724959650

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In this thesis ―Purification, Characterization and Binding of Metals to Zinc Alpha 2-Glycoprotein (ZAG) by Employing Various Techniques‖ the development of several techniques were carried out to analyze the interaction of biologically important metals of Irving William Series. The expression and purification of the ZAG protein was also optimized for the preparation of samples. The folding to the native conformation was also carried out. That ZAG protein is present in almost all fluids of the body. Since the discovery of ZAG, several studies have been carried out. The importance of ZAG as an important biochemical agent is known long before. The growing number of literature since today demonstrates that it’s an important biomarker for the cancer cachexia, prostate cancer, lipid mobilizing agent and obesity suppressor. The crystal structure of ZAG reveals that the fold and distribution of domains are similar to the Major histocompatibility complex-I (MHC-I) molecules. This class is known for their immune regulation and thus ZAG is believed as a central player in the immune system and cellular signaling. There is no clear evidence of ZAG participation in the cancerous conditions but over expression is dealt as a biomarker. The study shows that the ZAG interacts with several ligands. There interaction with metals is known for boosting the bioactivity of ZAG. Although, it’s believed long ago that the ZAG is an important protein that participate in several biochemical processes and little is known to understand the ligands bound conformations of ZAG with their targets like peptides and metals. In this current project, we were focused to get the good expression and purification procedure for the preparation of ZAG. The unnative aggregated fold of ZAG from the inclusion bodies was transformed into the native fold using refolding and dialysis method optimization. The introduction of histidine tag to the C-terminal and their successful removal by the Carboxypeptidase Y enzyme was also carried out. The results of the cleavage of histidine tag were evaluated with gel electrophoresis, Mass Spectrometry and Fluorescence Spectroscopy. The confirmation of native fold and bioactivity was also confirmed from DAUDA binding affinity using fluorescence emission. The formation of metal complex of Irving William Series with ZAG and their respective complexation analysis was also studies using MALDI-TOF MS and Fluorescence Spectroscopy. xiii It was found that the ZAG has multiple binding sites that interact with metals even in the presence of DAUDA. The X-ray tube analysis using FPXRF shows that Fe (III) and Co (II) has similar concentration of atoms bound to ZAG molecules.
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مولوی عزیز الرحمن

مولوی عزیز الرحمن
مولوی عزیز الرحمن صاحب کوئر یاپار اعظم گڑھ کے ایک شریف و نجیب خاندان سے تعلق رکھتے تھے، اردو کے مشہور ادیب و نقاد جناب شمس الرحمن فاروقی ان کے حقیقی بھتیجے تھے، جو اور اس خاندان کے دوسرے اشخاص بھی بڑے سرکاری عہدوں پر فائر ہیں۔ علمی و دنیاوی وجاہت کی طرح دینداری میں بھی یہ خانوادہ ممتاز تھا۔
مولوی عزیز الرحمن صاحب کی تعلیم مدرسہ الٰہیات کانپور میں ہوئی تھی اور انھوں نے الٰہ آباد بورڈ کے امتحانات بھی اچھے نمبروں سے پاس کئے تھے، ۱۹۲۵؁ء میں وہ شبلی نیشنل ہائر سکنڈری اسکول میں تدریس کی خدمت پر مامور ہوئے اور ۶۶؁ء میں ریٹائر ہوئے۔
مولوی صاحب کو قومی و ملی اشغال سے بھی سروکار رہا اور جمعیۃ علمائے ہند اور کانگریس پارٹی سے وابستہ رہے، اعظم گڑھ کے نسواں اسکول کے جواب گریجویٹ کالج ہوگیا ہے، بانی ارکان میں تھے، برسوں اس کے صدر بھی رہے۔
ملازمت کے ابتدائی زمانے سے دارالمصنفین آنے کا معمول بنالیا تھا۔ اس وضع داری کو اس وقت تک نباہا جب تک پیروں میں قوت رہی، انہیں مولانا سید سلیمان ندوی صاحبؒ اور مولوی مسعود علی ندوی کی مجلس میں باریاب ہونے کا شرف حاصل تھا، شاہ معین الدین احمد اور سید صباح الدین عبدالرحمن صاحبان اور دوسرے رفقا اور کارکنوں سے نہایت بے تکلف تھے، اس ناچیز پر بھی بہت شفقت فرماتے تھے۔
دو تین برس سے بالکل معذور اور خانہ نشین ہوگئے تھے، بالاخر ۲۸؍ اور ۲۹؍ دسمبر کی درمیانی شب میں واصل بحق ہوگئے، اﷲ ان کے درجات بلند کرے اور پسماندگان کر صبر جمیل عطا فرمائے، آمین۔ (ضیاء الدین اصلاحی۔ جنوری ۱۹۹۴ء)

 

Concept of Tolerance in Islam for Promotion of Human Unity

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Heat and Mass Transfer Enhancement in Rotating Disk Systems

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