Identification of N-terminus peptide of human granulocyte/macrophage colony stimulating factor as the site of nucleotide interaction.
The interaction of nucleotides with recombinant human granulocyte/macrophage colony stimulating factor (rhGM-CSF) has been investigated. Utilizing nucleotide photoaffinity probes [gamma 32P]-8N3ATP and [beta 32P]-8N3Ap4A, an analog of alarmone, the specificity of interaction was demonstrated by saturation of photoinsertion by these analogs and protection of photoinsertion by these analogs in the presence of natural nucleotide. The site of photoinsertion was tentatively determined to be Ser9. The photolabeled cytokine has lost most of its biological activity in a cellular proliferation assay, indicating a possible physiological role for this interaction.
Chavan
A J
AJ
College of Pharmacy, VA Medical Center, Lucille P. Markey Cancer Center University of Kentucky 40536.
Gass
C
C
Haley
B E
BE
Boone
T
T
Doukas
M A
MA
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D001386:Azides / Q000378:metabolism*; D001665:Binding Sites; D001681:Biological Assay; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
C092490:8-azidodiadenosine tetraphosphate; D001386:Azides; D015226:Dinucleoside Phosphates; D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D011994:Recombinant Proteins; C017040:8-azidoadenosine 5'-triphosphate; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000255:Adenosine Triphosphate