Chemical synthesis of yeast mitochondrial ATP synthase membranous subunit 8.
Chemical synthesis of highly hydrophobic peptides and proteins remains a challenging problem. Strong interchain associations within the peptide-resin matrix have to be overcome. A synthetic strategy for solid phase peptide synthesis is proposed, mainly based on prolonged coupling time using aprotic polar solvent mixtures. A tailored chromatographic purification was required to obtain a sample sufficiently pure for structural analysis. In this work, the total chemical synthesis of the membrane-embedded yeast mitochondrial ATP synthase subunit 8 is described. The quality of the synthetic protein was checked by electrospray mass spectrometry, its tendency to adopt alpha-helical secondary structure is evidenced by circular dichroism spectroscopy.
Goetz
M
M
Institut Europeén de Chimie et de Biologie, Ecole Polytechnique, Université Bordeaux I, Talence, France. michael.goetz@iecb-polytechnique.u-bordeaux.fr
Schmitter
J M
JM
Geoffre
S
S
Dufourc
E J
EJ
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D007425:Intracellular Membranes / Q000201:enzymology*; D008565:Membrane Proteins / Q000138:chemical synthesis; D008928:Mitochondria / Q000648:ultrastructure*; D017433:Protein Structure, Secondary; D006180:Proton-Translocating ATPases / Q000138:chemical synthesis*; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure; D015003:Yeasts / Q000648:ultrastructure
D008565:Membrane Proteins; D006180:Proton-Translocating ATPases