Yv. Griko et Md. Kapanadze, PURIFICATION AND CHARACTERIZATION OF HUMAN PANCREATIC-POLYPEPTIDE EXPRESSED IN ESCHERICHIA-COLI, Biochemical and biophysical research communications, 213(1), 1995, pp. 239-248
The region of cDNA encoding human pancreatic polypeptide (hPP) was obt
ained by polymerase chain reaction (PCR) and subcloned into an express
ion vector. The pancreatic polypeptide gene was expressed in Escherich
ia coli in two versions: as a cleavable fusion protein with IgG-bindin
g synthetic ZZ domains of protein A from Staphylococcus aureus or with
the 1-48 fragment of lambda Cro repressor, Site-specific hydrolysis b
y hydroxylamine was used to cleave the fusion protein, releasing the h
uman polypeptide. The structure of the obtained hPP has been studied b
y scanning microcalorimetry and circular dichroism spectrometry. It ha
s been shown that hPP in solutions close to neutral has a compact and
unique spatial structure with an extended hydrophobic core. This struc
ture is stable at 20 degrees C and co-operatively breaks down upon hea
ting from this temperature. (C) 1995 Academic Press, Inc.