Microwave Assisted Peptide Synthesis as a New Gold Standard in Solid Phase Peptide Synthesis: Phospholamban as an Example

Shadi Abu-Baker, Philip Garber, Bryce Hina, Trevor Reed, Ghaffari Shahrokh, Mohannad Al-Saghir, Gary Lorigan

College of Arts and Sciences, Ohio University, Zanesville, USA.
College of Arts and Sciences, Ohio University, Zanesville, USA; Genesis Hospital, Ohio University, Zanesville, USA.
Department of Chemistry and Biochemistry, Miami University, Oxford, USA.

DOI: 10.4236/ojsta.2014.31001                

Abstract

In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotection reactions during peptide synthesis. The use of microwave energy for both the coupling and deprotection steps makes the microwave peptide synthesizers the most versatile and powerful systems available. It produces high yield and fast synthesis when compared to conventional peptide synthesizers.

Keywords

Solid-State Peptide Synthesis; Phospholamban; Microwave Assisted Synthesis

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	H. K. B. Simmerman and L. R. Jones, “Phospholamban: Protein Structure, Mechanism of Action, and Role in Cardiac Function,” Physiological Reviews, Vol. 78, 1998, pp. 921-947.
[2]	P. James, M. Inui, M. Tada, M. Chiesi and E. Carafoli, “Nature and Site of Phospholamban Regulation of the Calcium Pump of Sarcoplasmic Reticulum,” Nature, Vol. 342, 1989, pp. 90-92. http://dx.doi.org/10.1038/342090a0
[3]	M. A. Kirchberger, M. Tada and A. M. Katz, “Phospholamban a Regulatory Protein of the Cardiac Sarcoplasmic Reticulum,” Recent Advances in Studies on Cardiac Structure and Metabolism, Vol. 5, 1975, pp. 103-115.
[4]	J. Fuji, A. Ueno, K. Kitano, S. Tanaka, M. Kadoma and M. Tada, “Complete Complementary DNA-Derived Amino Acid Sequence of Canine Cardiac Phospholamban,” Journal of Clinical Investigation, Vol. 79, 1987, pp. 301-304. http://dx.doi.org/10.1172/JCI112799
[5]	Q. Yao, J. L. Bevan, R. F. Weaver and D. J. Bigelow, “Purification of Porcine Phospholamban Expressed in Escherichia coli,” Protein Expression and Purification, Vol. 8, 1996, pp. 463-468. http://dx.doi.org/10.1006/prep.1996.0125
[6]	J. G. Collins, E. G. Kranias, A. S. Reeves, L. M. Bilezikjian and A. Schwartz, “Isolation of Phospholamban and a Second Proteolipid Component from Canine Cardiac Sarcoplasmic Reticulum,” Biochemical and Biophysical Research Communications, Vol. 99, 1981, pp. 796-803. http://dx.doi.org/10.1016/0006-291X(81)91235-3
[7]	E. J. Mayer, E. McKenna, V. M. Garsky, C. J. Burke, H. Mach, C. R. Middaugh, M. Sardana, J. S. Smith and R. G. Johnson Jr., “Biochemical and Biophysical Comparison of Native and Chemically Synthesized Phospholamban and a Monomeric Phospholamban,” The Journal of Biological Chemistry, Vol. 271, 1996, pp. 1669-1677. http://dx.doi.org/10.1074/jbc.271.3.1669
[8]	D. J. Hirsh, J. Hammer, W. L. Maloy, J. Blazyk and J. Schaefer, “Secondary Structure and Location of a Magainin Analogue in Synthetic Phospholipid Bilayers,” Biochemistry, Vol. 35, 1996, pp. 12733-12741. http://dx.doi.org/10.1021/bi961468a
[9]	A. Mascioni, C. Karim, J. Zamoon, D. D. Thomas and G. Veglia, “Solid-state NMR and rigid body molecular dynamics to determine domain orientations of monomeric phospholamban,” Journal of the American Chemical Society, Vol. 124, 2002, pp. 9392-9393. http://dx.doi.org/10.1021/ja026507m
[10]	E. K. Tiburu, P. C. Dave, K. Damodaran and G. A. Lorigan, “Investigating Leucine Side-Chain Dynamics and Backbone Conformations of Phospholamban Incorporated in Phospholipid Bilayers Utilizing 2H and 15N SolidState NMR Spectroscopy,” Biochemistry, Vol. 43, 2004, pp. 13899-13909. http://dx.doi.org/10.1021/bi0490993
[11]	http://brf.jcs.anu.edu.au/services/Peptidesynthesis/spps.html
[12]	“Derivatives for Enhanced Peptide Synthesis,” Novabiochem 2004/2005 Catalog, pp 185-188.
[13]	E. K. Tiburu, P. C. Dave, J. F. Vanlerberghe, T. B. Cardon, R. E. Minto and G. A. Lorigan, “An Improved Synthetic and Purification Procedure for the Hydrophobic Segment of the Transmembrane Peptide Phospholamban,” Analytical Biochemistry, Vol. 318, 2003, pp. 146-151. http://dx.doi.org/10.1016/S0003-2697(03)00141-6