عنوان

Implementing Mass Spectrometry for the Structural and Compositional Analysis of Proteins

شماره

TLpq2312810307

زبان متن نوشتاري يا گفتاري و مانند آن

انگلیسی

عنوان اصلي

Implementing Mass Spectrometry for the Structural and Compositional Analysis of Proteins

نام عام مواد

[Thesis]

نام نخستين پديدآور

Riaz, Mohammad

نام ساير پديدآوران

Sharp, Joshua S.

نام ناشر، پخش کننده و غيره

The University of Mississippi

تاریخ نشرو بخش و غیره

2019

نام خاص و کميت اثر

105

جزئيات پايان نامه و نوع درجه آن

M.S.

کسي که مدرک را اعطا کرده

The University of Mississippi

امتياز متن

2019

متن يادداشت

To understand the functions of proteins at a molecular level, it is often necessary to determine their three-dimensional structure. Fast Photochemical Oxidation of Protein (FPOP) is a hydroxyl-radical-based protein footprinting (HRPF) technique that utilizes a pulsed KrF laser (248 nm) to trigger photolysis of hydrogen peroxide to produce hydroxyl radicals, which subsequently modify the solvent exposed surface area of proteins. However, this technique has some disadvantages: being time-consuming, especially when dealing with a large sample size, adjustment of flow rate and difficulty with membrane protein oxidation. To address these issues, we developed a platform to perform FPOP in microtiter plates instead of the traditional capillary set up. To ensure reliability and reproducibility of microtiter FPOP and evaluate microtiter FPOP against traditional flow FPOP, we used three systems: adenine-based hydroxyl radical dosimetry; oxidation of the model peptide [Glu]1-Fibrinopeptide B (GluB); and HRPF analysis of the model protein myoglobin. We demonstrated that microtiter based FPOP can provide comparable oxidation of model peptide and model protein as compared to traditional capillary FPOP. Automation of the system substantially reduces experimental time and minimize human errors. Lectins are sugar-binding proteins that perform various biological functions such as cellular recognition, attachments, etc. Pulses, the dried edible seeds of certain plants, are great sources of lectins, and a proteomics approach directed at lectins identification would bring substantial improvement in the lectin identification process. Lectins from nine different plant species were analyzed through SDS-PAGE and proteomics analysis. After LC-MS/MS analysis, proteins were identified with protein database searches using Uniprot. Functionally uncharacterized proteins were identified with database searches, were annotated with the Pfam and NCBI-CCD databases. In-vitro pharmacological screening will be carried out to assess the pharmacological effects of these lectins.

موضوع مستند نشده

Pharmaceutical sciences

موضوع مستند نشده

Pharmacology

مستند نام اشخاص تاييد نشده

Riaz, Mohammad

مستند نام اشخاص تاييد نشده

Sharp, Joshua S.

نام الکترونيکي

فرمت انتشار

p

نوع ماده

[Thesis]

کد کاربرگه

276903

سطح دسترسي

a

تكميل شده

Y