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Investigation of Active Site of Osrglpi. a Germin-Like Superoxide Dismuatse

Thesis Info

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Author

Dur E Shahwar, ,

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Chemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/13455/1/Shahwar%20PhD%20Thesis%20final%20final.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726500841

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Germin-like proteins are glycosylated proteins that are highly diverse in plant kingdom. Most of the germin-like proteins show superoxide dismutase activity. Superoxide dismutase converts superoxide anion into molecular oxygen and hydrogen peroxide. Identification of catalytic residues is an important step for understanding the mechanism of enzyme catalyzed reactions. Previously OsRGLP1 gene and cDNA were isolated from the roots of Oryza sativa and, under CaMV35S promoter, cDNA has been heterologously expressed in tobacco. In transgenic tobacco plants, OsRGLP1 seems to show SOD activity. Present study was aimed to determine the active site of OsRGLP1. Three dimensional structure and residues involved in metal ion and substrate binding were predicted using bioinformatics tools. Three histidines and one glutamate were predicted to play an important role in metal ion binding and Asparagine to Alanine mutation was predicted to be responsible for different substrate specificity of germin and germin-like proteins. These residues were then mutated with other selected residues by site directed mutagenesis to confirm these predictions. In present study out of many designed mutants, four loss of function mutants were further studied by performing transient transformation of tobacco plants. These transgenic plants were used as source of native and mutant proteins for SOD activity assay to find the effect of mutations on SOD activity. Results of SOD assay showed approximately complete loss of activity in all mutant proteins validating the importance of these residues in SOD activity of OsRGLP1 protein. As these residues are involved in metal ion binding so present study also provide some insight into mechanism of action of OsRGLP1 indicating OsRGLP1 indicating the role of metal ion in its activity.
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مولوی محبوب عالم

مولوی محبوب عالم
اس ماہ کے شذرات کا صفحہ وفات نامہ ہوا چاہتا ہے، مگر احسان فراموشی ہوگی اگر ملک کے سب سے بوڑھے صحیفۂ نگار مولوی محبوب عالم اڈیٹر پیسہ اخبار لاہور کا ماتم نہ کیا جائے، ۲۸؍ مئی کو انہوں نے اس دارِ فانی کو الوداع کہا، وہ اردو کے سب سے پہلے روزنامہ اخبار (پیسہ) کے اڈیٹر تھے، انہوں نے صرف اپنی محنت و کوشش سے سرمایہ حاصل کیا اور ملک میں تاریخ اور سیاحت ناموں کے پڑھنے کا ذوق پیدا کیا اور خود بھی یورپ اور ممالک اسلامیہ کے دوسفر کئے اور سیاحت نامے لکھے، مگر افسوس کہ اب ان کو وہ سفر پیش آیا جس کا سفرنامہ انسانوں کے ہاتھ نہیں، فرشتوں کے ہاتھ لکھتے ہیں، اس ان دیکھی منزل کے بوڑھے مسافر پر اﷲ تعالیٰ کی رحمت ہو۔
مرحوم نے ۷۴ برس کی عمر پائی۔ (سید سلیمان ندوی، جولائی ۱۹۳۳ء)
تصحیح: منشی محبوب عالم مرحوم کے تذکرہ میں یہ لکھا گیا ہے کہ وہ اردو کے پہلے روزانہ اخبار کے بانی اور اڈیٹر تھے، اس سے مراد مسلمانوں میں تھی، یعنی اردو کے پہلے اسلامی روزانہ اخبار کے وہ بانی اور اڈیٹر تھے، اردو میں منشی نولکشور لکھنو کا اودھ اخبار ان کے اخبار سے پہلے نکلا تھا اور اب تک نکل رہا ہے۔ (سید سلیمان ندوی، اگست ۱۹۳۳ء)

 

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