Synthesis of the pro-peptide of subtilisin BPN'.
Subtilisin, a bacterial serine protease, is secreted as pre-pro-subtilisin. Previously, we demonstrated that the pro-peptide moiety of intact pro-subtilisin can guide the folding of inactive protein to active enzyme both in an intramolecular (6) and intermolecular manner (18). Herein is reported the total chemical synthesis of the pro-sequence (77 amino acids) of pre-pro-subtilisin BPN' carried out by solid phase methods. The structure was confirmed by both sequencing and amino acid analysis of the fragment peptides resulting from a V-8 protease digest. Preliminary studies indicate that the synthetic pro-peptide itself can renature denatured subtilisin BPN'. This study demonstrates a novel method for examining protein folding with the aid of exogenously added synthetic peptides.
Cash
P W
PW
Rutgers University, Newark, NJ 07102.
Zhu
X
X
Ohta
Y
Y
Tsao
J
J
Lackland
H
H
Mateos-Nevado
M D
MD
Inouye
M
M
Stein
S
S
Jordan
F
F
Tous
G I
GI
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D004792:Enzyme Precursors / Q000138:chemical synthesis* / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000378:metabolism; D010449:Peptide Mapping; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D012697:Serine Endopeptidases; D013381:Subtilisins / Q000138:chemical synthesis* / Q000378:metabolism
D004792:Enzyme Precursors; D010446:Peptide Fragments; D010455:Peptides; C057107:prosubtilisin; D012697:Serine Endopeptidases; D013381:Subtilisins; C028344:glutamyl endopeptidase; D003488:Cyanogen Bromide