Combined solid-phase/solution synthesis of large ribonuclease A C-terminal peptides containing a non-natural proline analog.
Three large peptides corresponding to the 65-124 (60-mer), 72-124 (53-mer), and 77-124 (48-mer) sequence of bovine pancreatic ribonuclease A (RNase A) were assembled from either two or three shorter protected peptide fragments by chemical coupling in solution. The fragments were synthesized manually by 9-fluorenylmethyloxycarbonyl (Fmoc)-based solid-phase peptide chemistry in plastic syringes, and subsequently purified by normal-phase high-performance liquid chromatography on a silica gel column. The main aim of this work was to incorporate sterically hindered l-5,5-dimethylproline (dmP) as a substitute for Pro(93) into the sequence of RNase A in order to constrain the -Tyr(92)-Pro(93)- peptide group to a single cis-conformation.
Cerovský
V
V
Baker Laboratory of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853-1301, USA.
Scheraga
H A
HA
D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000235:genetics / Q000302:isolation & purification; D011392:Proline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010446:Peptide Fragments; D011392:Proline; D012259:Ribonuclease, Pancreatic; C003342:stachydrine