Characterization and purification of the Drosophila Kc cell JH binding protein

Thesis/Dissertation · Sun Jan 01 04:00:00 EST 1989

OSTI ID:6159584

Wang, Xiuhua

Juvenile hormone (JH) is one of the principal regulators of insect development. First, physical properties of the JH binding protein are characterized using a photoaffinity ligand ({sup 3}H)-EFDA. Second, ({sup 3}H)-EFDA is employed as labeled to examined the behavior of the JH binding protein in several protein separation procedures. The properties of heparin-agarose, hydroxylapatite, gel filtration, ion exchange, and chromatofocusing chromatography are examined for their ability to affect separation of the JH binding protein from other components of Kc cell cytosol. Third, several of the separation procedures are linked sequentially into a JH binding protein purification scheme and the procedure is shown to purify the JH binding protein to homogeneity. Fourth, competition study is carried out to demonstrate the protein purified using ({sup 3}H)-EFDA is JH competable. In addition, JH III, the naturally occurring hormone, is used as the radioligand in this purification procedure, the authenticity of the intracellar JH binding protein is reassured. Fifth, the amino acid composition of the JH binding protein is elucidated. Finally, nanomoles of the purified JH binding protein are collected for protein microsequencing. The sequencing data will be used to synthesize peptide which, in turn, will be used for JH binding protein antibody production.

• Research Organization: California Univ., Los Angeles, CA (USA)
• OSTI ID: 6159584
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANIMALS
ARTHROPODS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
DIPTERA
DROSOPHILA
FLIES
FRACTIONATION
FRUIT FLIES
HORMONES
HYDROGEN COMPOUNDS
INSECTS
INVERTEBRATES
ISOTOPE APPLICATIONS
LIGANDS
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
PHYSICAL PROPERTIES
PROTEINS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRITIUM COMPOUNDS