Synthetic Method
The RNase 1-118 was prepared by successive digestion of RNase A pepsin and
carboxypeptidase A (Doscher, 1983). The synthetic component, RNase 111-124,
was prepared by the use of solid-phase peptide synthetic mothods, in which
the peptide chain was assembled in the stepwise mannar while it was attached
at one end to a solid support. The peptide chain was extented by repetitive
steps of deprotection, neutralization and coupling until the desired sequence
was obtained (Lin, 1970). It was important that the synthesis proceeds rapidly
and in high yields to prevent side reactions or by-products.
Related Web-links
1. Introduction to Ribonuclease A by Raines:
http://www.uta.edu/faculty/sawasthi/Enzymology-4351-5324/Class%20Syllabus%20Enzymology/ribonucleaseA.pdf
2. Solid Phase Synthesis by Merrifield (Nobel Prize Winner):
http://nobelprize.org/nobel_prizes/chemistry/laureates/1984/merrifield-lecture.pdf
3. Chemical Synthesis of Proteins:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2845543/?tool=pmcentrez
4. Refined Crystal Structure: http://www.ncbi.nlm.nih.gov/pubmed/3680234
References
Martin, Philip D., Marilynn S. Doscher, and Brian F. P. Edwards. "The Redefined
Crystal Structure of a Fully Active Semisynthetic Ribonuclease at 1.8-A Resolution."
The Journal of Biological Chemistry
262.33 (1987): 15930-5938.
Marilynn S. Doscher, Philip D. Martin and Brian F.P. Edwards, "Characerization
of the Histidine Proton Nuclear Magnetic Resonance of a Semisynthetic Ribonuclease."
Biochemistry, 1983,22,4125-4131.
Lin, M. C. (1970) Journal of Biological Chemistry, 245, 6726-6731.
David J. Boerema, Valentina. A. T., Stephen B. H. Kent, "Total Synthesis by
Modern chemical Ligation Methods and High Resolution (1.1-A) X-ray structure
of Ribonuclease A. Biopolymers. 2008;90(3):278-86.
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