Structural mapping of post-translational modifications in human interleukin-24: role of N-linked glycosylation and disulfide bonds in secretion and activity.
Human interleukin-24 (IL-24) is unique among the IL-10 superfamily as there is considerable evidence that it possesses multiple anti-cancer properties, including direct tumor cell cytotoxicity, helper T cell (TH1) immune stimulation, and anti-angiogenic activities. The primary sequence of human IL-24 differs from homologous cytokines, because it possesses three consensus N-linked glycosylation sites and the potential for a single disulfide bond. To address the significance of these modifications in human IL-24, we analyzed the relationship between post-translational modifications and the cytokine activity of the human IL-24 protein. In contrast to related interleukins, we identified a relationship between net glycosylation, protein solubility, and cytokine activity. In addition, abrogation of the two cysteine residues by mutagenesis dramatically altered the ability of IL-24 to secrete from host cells and resulted in the concomitant loss of IL-24 activity. We conclude that, unlike other IL-10 family members, human IL-24 must be glycosylated to maintain solubility and bioavailability. Further, a single, unique disulfide bond is required for secretion and activity. These structure-function relationships show that, although IL-24 is a member of the IL-19 subfamily of IL-10-like cytokines by sequence similarity, its surface properties and its distinctive disulfide arrangement make it unique. These observations could explain the novel biological activities measured of this cytokine. Understanding the structural basis of IL-24 activity will be important in the interpretation of the function of this cytokine and in the development of scale-up strategies for biophysical and clinical applications.
Fuson
Kerry L
KL
Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, USA.
Zheng
Mingzhong
M
Craxton
Molly
M
Pataer
Abujiang
A
Ramesh
Rajagopal
R
Chada
Sunil
S
Sutton
R Bryan
RB
D003545:Cysteine / Q000235:genetics; D016207:Cytokines; D004220:Disulfides; D006031:Glycosylation; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D012995:Solubility; D013329:Structure-Activity Relationship; D000099180:Interleukin-24
D003545:Cysteine; D016207:Cytokines; D004220:Disulfides; D007378:Interleukins; D000099180:Interleukin-24