Studi Proteomik pada Protein Otot Rangka Rattus norvegicus Menggunakan Spektrometri Massa Resolusi Tinggi

Autor(s): Alvina Nur Aini
DOI: 10.55075/wa.v46i1.97

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Analisis proteomik bertujuan untuk memperoleh informasi mengenai ekspresi protein seluler. Oleh karena fungsi penting protein dalam organisme hidup, proteomik memainkan peran penting dalam memahami sistem dan proses biologi. Metode paling umum untuk identifikasi protein dalam proteomik adalah menggunakan Mass Spectrometer (MS). Secara lebih spesifik, instrumen MS yang digunakan dalam analisis proteomik adalah Spektrometri Massa Resolusi Tinggi, yang mampu menganalisis massa eksak suatu ion molekuler. Dalam penelitian ini dilakukan hidrolisis sampel protein otot rangka Rattus norvegicus dengan menggunakan enzim tripsin untuk mendapatkan gambaran mengenai proses analisis proteomik. Hidrolisis protein dilakukan dengan variasi enzim:substrat 1:25 (b/b). Hasil hidrolisis kemudian diidentifikasi dengan menggunakan Spektrometri Massa Resolusi Tinggi. Berdasarkan hasil analisis, terdapat 219 protein yang terindentifikasi. Hasil fragmentasi setiap peptida protein menunjukkan kesesuaian dengan fragmen teoritis secara in silico. Hal ini mengindikasikan bahwa identifikasi peptida menggunakan Spektrometri Massa Resolusi Tinggi mampu memberikan hasil yang akurat, sehingga dapat menghindari keraguan dalam identifikasi protein yang kompleks. Hasil ini juga menunjukkan bahwa Spektrometri Massa Resolusi Tinggi memungkinkan screening target peptida dan identifikasi peptida dari matriks protein seluler.

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Referensi

Alfiraza, E.N., 2018, Identifikasi Peptida Spesifik pada Gelatin Babi Menggunakan Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS), Tesis, Universitas Gadjah Mada, Yogyakarta.

Domon, B., & Aebersold, R. (2006). Mass Spectrometry and Protein Analysis, Sciences, 312(5771), 212-217.

Gasteiger, E., Gattiker, A., Hoogland, C., Ivanyi, I., Appel, R.D., & Bairoch, A. (2003). ExPASy: The Proteomics Server for In-Depth Protein Knowledge and Analysis, Nucleic Acids Research, 31, 3784-3788.

http://db.systemsbiology.net/, diakses pada 20 Mei 2022.

Hustoft, H.K., Malerod, H., Wilson, S.R., Reubsaet, L., Lundanes, E. and Greibrokk, T., 2012, A Critical Review of Trypsin Digestion for LC-MS Based Proteomics, Integrative Proteomics, Rijeka: InTech.

Kapteyn, J.C., Saidi, M.D., Dijkstra, R., Kars, C., Tjon, J.C., Weverling, G.J., de Vocht, M. L., Kompier, R., van Montfort, B. A., Guichoux, J-Y., Goudsmit, J., & Lagerwerf, F. M. (2006). Haemagglutinin Quantification and Identification of Influenza A&B Strains Propagated in PER.C6(R) Cells: A Novel RP–HPLC Method, Vaccine, 24, 3137–3144.

Kenyon, G.L., DeMarini, D.M., Fuchs, E., Galas, D.J., Kirsch, J.F., Leyh, T.S., Moos, W.H., Petsko, G.A., Ringe, D., Rubin, G.M., & Sheahan, L.C. (2002). Defining the Mandate of Proteomics in the Post-Genomics Era: Workshop Report, Molecular & Cellular Proteomics, 1(10), 763–780.

McLafferty, F.W., Breuker, K., Jin, M., Han, X., Jiang, H., Kong, X., & Begley, T.P. (2007). Top-Down MS: a Forward Complement to the High Capabilities of Proteolysis Proteomics, FEBS Journal, 274, 6256-6268.

Moriwaki, K.T., & Shiroishi, H.Y. (1994). Genetic In Wild Mice, Its Application to Biomedical Research, Tokyo: Karger.

Orduna, A.R., Husby, E., Yang, C.T., Ghosh, D. and Beaudy, F., 2017, Detection of Meat Species Adulteration Using High-Resolution Mass Spectromery and a Proteogenomics Strategy, Food Additives & Contaminats, 34(7).

Ortrea, I., O’Connor, G., & Maquet, A. (2016). Review on Proteomics for Food Authentication, Proteomics, 147, 212-225.

Pan, S., Zhang, H., Rush, J., Eng, J., Zhang, N., & Patterson, D. (2005). High-Throughput Proteome-Screening for Biomarker Detection, Molecular & Cellular Proteomics, 4(2), 182-190.

Pandey, A., & Mann, M. (2000) Proteomics to Study Genes and Genomes, Nature, 405, 837-846.

Pavia, D.L., Lampman, G.M., Kriz, G.S., & Vyvyan, J.A. (2009). Introduction to Spectroscopy, 4th Ed., Boston: Cengage Learning.

Rebecchi, K.R., Go, E.P., Xu, L., Woodin, C.L., Mure, M., & Desaire, H. (2011). A General Protease Digestion Procedure for Optimal Protein Sequence Coverage and PTM Analysis of Recombinant Glycoproteins: Application to the Characterization of HLOXL2 Glycosylation, Analytical Chemistry, 83(22), 8484-8491.

Silva, J.C., Gorenstein, M.V., Li, G-Z., Vissers, J.P.C., & Geromanos, S.J. (2006). Absolute Quantification of Proteins by LCMSE: A Virtue of Parallel MS Acquisition, Molecular & Cellular Proteomics, 5, 144–156.

Smith, B.J. (2002). Protein Sequencing Protocols, Totowa: Humana.

Steen, H., & Mann, M. (2004). The ABC’s (and XYZ’s) of Peptide Sequencing, Molecular & Cellular Biology, 5, 699-711.

Suder, P., Bierczynska, A., Konig, S., & Silberring, J. (2004). Acid-Labile Surfactant Assists In-Solution Digestion of Proteins Resistant to Enzymatic Attack, Rapid Communication Mass Spectrom, 18 (2004), 822–824.

Weisser, H. and Choudhary, J.S., 2017, Targeted Feature Detection for Data-Dependent Shotgun Proteomics, J. Proteome Research, 16(8), 2964-2974.

Wierenga, S.K., Zocher, M.J., Mirus, M.M., Conrads, T.P., Goshe, M.B., & Veenstra, T.D. (2002). A Method to Evaluate Tryptic Digestion Efficiency for High-Throughput Proteome Analyses, Rapid Communication Mass Spectrom, 16, 1404–1408.

Wilkins, M.R., Pasquali, C., Appel, R.D., Ou, K., Golaz, O., & Sanchez, J.C. (1996). From Proteins to Proteomes: Large Scale Protein Identification by Two-Dimentional Electrophoresis and Amino Acid Analysis, Biotechnology, 14(1), 61-65.

DOI: https://doi.org/10.55075/wa.v46i1.97

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