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Expression and structure of the human NGF receptor

Thesis/Dissertation ·
OSTI ID:5516646
Nerve growth factor (NGF) plays an essential role in the development and maintenance of sensory and sympathetic neurons of the peripheral nervous system. Recent evidence indicates that NGF also acts in the central nervous system. We have used fragments of the human NGF receptor gene to isolate human NGF receptor cDNA clones. The cDNA clones have been sequenced, and the organization of the NGF receptor mRNA as well as the primary structure of the receptor protein have been determined. The message contains a short 5{prime} nontranslated region and a long 3{prime} nontranslated region. The receptor protein contains 399 amino acids, is cysteine-rich, and spans the membrane once. A full-length NGF receptor cDNA was cloned into a mammalian expression vector and transfected into NGFR-mouse L cells. Binding of {sup 125}I-NGF to transfectant cell lines was specific and saturable, verifying that the cloned protein represents an authentic NGF receptor. Transfectant cell lines overexpressing the human receptor were used to immunize syngenic animals and polyclonal antisera specific for the human NGF receptor were isolated. A series of NGF receptor mutants with truncated carboxyl termini were constructed and introduced into mouse L cells. Mutant proteins which contain intact transmembrane domains retain NGF binding activity and are localized to the plasma membrane.
Research Organization:
Princeton Univ., NJ (USA)
OSTI ID:
5516646
Country of Publication:
United States
Language:
English