Photoaffinity labeling of ATP and NAD sup + binding sites on recombinant human interleukin 2

Journal Article · Thu Feb 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.3.1243· OSTI ID:7064215

Campbell, S; Kim, H; Doukas, M; Haley, B ^[1]

---

1. Univ. of Kentucky, Lexington (USA)

Interleukin 2 (IL-2) is a T-cell-derived lymphokine critical in the activation and proliferation of T cells, B cells, and lymphokine-activated killer cells. It is a glycoprotein of {approx}15,500 daltons that is synthesized and secreted after activation by antigen or mitogen. By using the analogs 8-azidoadensoine 5{prime}-({gamma}-{sup 32}P)triphosphate (({gamma}-{sup 32}P)8N{sub 3}ATP) and nicotinamide 2-azidoadenine (adenylate-{sup 32}P)dinucleotide (({alpha}-{sup 32}P)2N{sub 3}NAD{sup +}) as photoaffinity probes, the authors have detected specific, metal ion-requiring nucleotide binding sites on recombinant human IL-2 (rhIL-2). The specificity of these nucleotide interactions with rhIL-2 was demonstrated by saturation effects and by competition by the parent nucleotides at physiologically relevant concentrations. Saturation of photoinsertion into rhIL-2 occurred at 50 {mu}M ({gamma}-{sup 32}P)8N{sub 3}ATP. Saturation of photoinsertion with ({alpha}-{sup 32}P)2N{sub 3}NAD{sup +} was observed at 180 {mu}M. The extent of photoinsertion of both photoprobes into rhIL-2 varied with the presence of different divalent metal ions. The biological significance of these interactions is unknown, but their specificity suggests that nucleotide binding may be involved in the bioactivity of IL-2.

• OSTI ID: 7064215
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:3; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METAL COMPOUNDS
ANIMAL CELLS
ANIMALS
ATP
AZIDO COMPOUNDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CALCIUM CHLORIDES
CALCIUM COMPOUNDS
CALCIUM HALIDES
CATIONS
CELL PROLIFERATION
CHARGED PARTICLES
CHEMICAL REACTIONS
CHLORIDES
CHLORINE COMPOUNDS
COENZYMES
CONNECTIVE TISSUE CELLS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
ELEMENTS
GLYCOPROTEINS
GROWTH FACTORS
HALIDES
HALOGEN COMPOUNDS
IONS
ISOTOPES
KINETICS
LEUKOCYTES
LIGHT NUCLEI
LYMPHOCYTES
LYMPHOKINES
MAGNESIUM CHLORIDES
MAGNESIUM COMPOUNDS
MAMMALS
MAN
MATERIALS
METALS
MITOGENS
NAD
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POLYMERIZATION
PRIMATES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
SOMATIC CELLS
VERTEBRATES
ZINC CHLORIDES
ZINC COMPOUNDS
ZINC HALIDES