Expression, purification and crystallization of human prolylcarboxypeptidase.
Prolylcarboxypeptidase (PrCP) is a lysosomal serine carboxypeptidase that cleaves a variety of C-terminal amino acids adjacent to proline and has been implicated in diseases such as hypertension and obesity. Here, the robust production, purification and crystallization of glycosylated human PrCP from stably transformed CHO cells is described. Purified PrCP yielded crystals belonging to space group R32, with unit-cell parameters a = b = 181.14, c = 240.13 A, that diffracted to better than 2.8 A resolution.
Abeywickrema
Pravien D
PD
Global Structural Biology, Merck Research Laboratories, West Point, PA 19486, USA.
Patel
Sangita B
SB
Byrne
Noel J
NJ
Diehl
Ronald E
RE
Hall
Dawn L
DL
Ford
Rachael E
RE
Rickert
Keith W
KW
Reid
John C
JC
Shipman
Jennifer M
JM
Geissler
Wayne M
WM
Pryor
Kelly D
KD
SinhaRoy
Ranabir
R
Soisson
Stephen M
SM
Lumb
Kevin J
KJ
Sharma
Sujata
S
D000818:Animals; D016466:CHO Cells; D002268:Carboxypeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D006224:Cricetinae; D003412:Cricetulus; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans
D002268:Carboxypeptidases; C022448:lysosomal Pro-X carboxypeptidase