» Articles » PMID: 3263644

Role of Disulfide Bridges in Determining the Biological Activity of Interleukin 3

Overview
Specialty Science
Date 1988 Nov 1
PMID 3263644
Citations 8
Authors
Affiliations
Soon will be listed here.
Abstract

Total chemical synthesis of analog proteins was used to examine the requirement for specific disulfide bridges for the biological activity of interleukin 3 (IL-3), a growth factor that stimulates multiple lineages of hemopoietic cells. Four structural analogs of the mature, 140 amino acid murine IL-3 molecule were synthesized in which specific cysteine residues were replaced by alanines. In a quantitative IL-3 assay, based on [3H]thymidine incorporation into factor-dependent cells, the IL-3 analog with alanines substituted for all four cysteines--i.e., [Ala17,79,80,140]IL-3--had 1/500th as much activity as the molecule synthesized according to the native sequence. The two analogs [Cys17,79,Ala80,140]IL-3 and [Cys17,140,Ala79,80]IL-3 had similarly low activity, whereas the [Cys17,80,Ala79,140]IL-3 analog had 2000-fold higher activity than these three analogs, and 3-fold higher than the molecule with the native sequence. This shows that in IL-3 a single disulfide bridge, between cysteines 17 and 80, is required for biological activity that approximates physiological levels. This disulfide probably stabilizes the tertiary structure of the protein to give a conformation that is optimal for function.

Citing Articles

Structural and evolutionary exploration of the IL-3 family and its alpha subunit receptors.

Fogha J, Bayry J, Diharce J, de Brevern A Amino Acids. 2021; 53(8):1211-1227.

PMID: 34196789 DOI: 10.1007/s00726-021-03026-3.


Molecular evolution of interleukin-3.

Burger H, Wagemaker G, Leunissen J, Dorssers L J Mol Evol. 1994; 39(3):255-67.

PMID: 7932787 DOI: 10.1007/BF00160149.


Effects of interleukin-3 with or without the c-kit ligand, stem cell factor, on the survival and cytoplasmic granule formation of mouse basophils and mast cells in vitro.

Dvorak A, Seder R, Paul W, Morgan E, Galli S Am J Pathol. 1994; 144(1):160-70.

PMID: 7507298 PMC: 1887107.


Neutrophil-activating properties of the melanoma growth-stimulatory activity.

Moser B, Clark-Lewis I, Zwahlen R, Baggiolini M J Exp Med. 1990; 171(5):1797-802.

PMID: 2185333 PMC: 2187876. DOI: 10.1084/jem.171.5.1797.


Enhanced hematopoietic activity of a human granulocyte/macrophage colony-stimulating factor-interleukin 3 fusion protein.

Curtis B, Williams D, Broxmeyer H, Dunn J, Farrah T, Jeffery E Proc Natl Acad Sci U S A. 1991; 88(13):5809-13.

PMID: 1829529 PMC: 51967. DOI: 10.1073/pnas.88.13.5809.


References
1.
Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

2.
Ellman G . Tissue sulfhydryl groups. Arch Biochem Biophys. 1959; 82(1):70-7. DOI: 10.1016/0003-9861(59)90090-6. View

3.
Clark-Lewis I, Schrader J . P cell-stimulating factor: biochemical characterization of a new T cell-derived factor. J Immunol. 1981; 127(5):1941-7. View

4.
Sarin V, Kent S, Tam J, MERRIFIELD R . Quantitative monitoring of solid-phase peptide synthesis by the ninhydrin reaction. Anal Biochem. 1981; 117(1):147-57. DOI: 10.1016/0003-2697(81)90704-1. View

5.
Iscove N, Roitsch C, Williams N, Guilbert L . Molecules stimulating early red cell, granulocyte, macrophage, and megakaryocyte precursors in culture: similarity in size, hydrophobicity, and charge. J Cell Physiol Suppl. 1982; 1:65-78. DOI: 10.1002/jcp.1041130412. View