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A synthetic analogue of the insect juvenile hormone (JH) III, 10,11-epoxy[10-3H]farnesyl diazoacetate [( 3H]-EFDA), binds to several proteins in a partially purified preparation of hemolymph protein from fourth instar larvae of Manduca sexta when irradiated with UV light. Approximately 80% of this binding could be inhibited by the addition of excess unlabeled JH I. To compare the relative affinity of EFDA for the juvenile hormone binding protein (JHBP) with that of the various JH homologues, the ability of unlabeled EFDA and JH homologues to displace [3H]JH I from binding sites was measured. The relative affinities were EFDA greater than JH I greater than JH II greater than JH III. When Scatchard analysis of the binding of [3H]EFDA or [3H]JH I to the larval JHBP was performed, an estimated apparent KD of 4.5 X 10(-8)M was found for EFDA, whereas for JH I a slightly higher KD of 8.8 X 10(-8) M was calculated. To determine if [3H]EFDA bound at the JH I binding site, displacement of [3H]JH I from the JHBP complex with unlabeled JH I, JH II, and JH III was compared to the displacement of [3H]EFDA with the same homologues. The results demonstrated that the photoaffinity label bound covalently at the JH I binding site on the hemolymph binding protein of Manduca sexta. Fluorescence autoradiography of [3H]EFDA photoaffinity labeled proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that [3H]EFDA bound covalently to two major proteins in the absence of JH I.(ABSTRACT TRUNCATED AT 250 WORDS)
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
October 2003,
Molecular and cellular endocrinology,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
February 1974,
Proceedings of the National Academy of Sciences of the United States of America,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
August 1993,
Biochemistry,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
August 2007,
Insect biochemistry and molecular biology,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
April 1993,
Archives of biochemistry and biophysics,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
November 1990,
The Journal of biological chemistry,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
May 1978,
General and comparative endocrinology,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
January 2003,
Insect biochemistry and molecular biology,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
January 1985,
Methods in enzymology,
J K Koeppe, and
G D Prestwich, and
J J Brown, and
W G Goodman, and
G E Kovalick, and
T Briers, and
M D Pak, and
L I Gilbert
May 1977,
Biochemistry,
