A Structure-function
Analysis of Human GDNF
CHEN Zhe-Yu1,2, HE Zhi-Yong2, HE Cheng1,
LU Chang-Lin1, WU Xiang-Fu2*
(1Department of Neurobiology, the Second Military Medical
University, Shanghai 200433, China; 2Shanghai Institute of
Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China )
Abstract The glial cell
line-derived neurotrophic factor(GDNF) plays a very important role in the
regeneration of the nervous system. Based on the results of the X-ray structure
analysis of rat GDNF, human GDNF gene was modified with
deletion and insertion mutagenesis by using PCR methods. The various mutants
were all highly expressed in E.coli. The recombinant proteins
were purified and their survival-promoting activities were determined by using
cultures of the spinal cord neurons of embryonic mouse. The results showed that
the “cystine knot motif” was critical for the maintenance of GDNF structure； the α-helix, finger 1 and finger 2
region were critical for GDNF neurotrophic activity； and the N-terminus of human GDNF was not
essential for its biological functions.
Key words GDNF； gene mutant； structure-function analysis
Received:
November 22, 1999 Accepted: December 16, 1999
This work was supported by State Key Program of Basic
Research(973)
* Corresponding author: Tel, 86-21-64374430-292; Fax,
86-21-64338357; e-mail, [email protected]