Structural analysis of the receptors for granulocyte colony-stimulating factor on neutrophils

Journal Article · Thu Nov 01 04:00:00 EST 1990 · Experimental Hematology (Lawrence, Kansas); (USA)

OSTI ID:6284086

Hanazono, Y; Hosoi, T; Kuwaki, T; Matsuki, S; Miyazono, K; Miyagawa, K; Takaku, F ^[1]

---

1. Univ. of Tokyo (Japan)

We investigated granulocyte colony-stimulating factor (G-CSF) receptors on neutrophils from three patients with chronic myelogenous leukemia (CML) in the chronic phase, in comparison with four normal volunteers. Because we experienced some difficulties in radioiodinating intact recombinant human G-CSF, we developed a new derivative of human G-CSF termed YPY-G-CSF. It was easy to iodinate this protein using the lactoperoxidase method because of two additional tyrosine residues, and its radioactivity was higher than that previously reported. The biological activity of YPY-G-CSF as G-CSF was fully retained. Scatchard analysis demonstrated that CML neutrophils had a single class of binding sites (1400 +/- 685/cell) with a dissociation constant (Kd) of 245 +/- 66 pM. The number of sites and Kd value of CML neutrophils were not significantly different from those of normal neutrophils (p greater than 0.9). Cross-linking studies revealed two specifically labeled bands of (125I)YPY-G-CSF-receptor complexes with apparent molecular masses of 160 and 110 kd on both normal and CML neutrophils. This is the first report describing two receptor proteins on neutrophils. According to the analyses of the proteolytic process of these cross-linked complexes and proteolytic mapping, we assume that alternative splicing or processing from a single gene may generate two distinct receptor proteins that bind specifically to G-CSF but have different fates in intracellular metabolism.

• OSTI ID: 6284086
• Journal Information: Experimental Hematology (Lawrence, Kansas); (USA), Journal Name: Experimental Hematology (Lawrence, Kansas); (USA) Vol. 18:10; ISSN EXHMA; ISSN 0301-472X
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
ELECTRON CAPTURE RADIOISOTOPES
GROWTH FACTORS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
MEMBRANE PROTEINS
MITOGENS
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PATIENTS
POLYMERIZATION
PROTEINS
PROTEOLYSIS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
TRACER TECHNIQUES