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.
Global Structural Biology, Merck Research Laboratories, West Point, PA 19486, USA.
Pravien D
PD
Abeywickrema
Sangita B
SB
Patel
Noel J
NJ
Byrne
Ronald E
RE
Diehl
Dawn L
DL
Hall
Rachael E
RE
Ford
Keith W
KW
Rickert
John C
JC
Reid
Jennifer M
JM
Shipman
Wayne M
WM
Geissler
Kelly D
KD
Pryor
Ranabir
R
SinhaRoy
Stephen M
SM
Soisson
Kevin J
KJ
Lumb
Sujata
S
Sharma
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