Disulfide assignments in recombinant mouse and human interleukin 4

Carr, Christina; Aykent, Serdar; Kimack, Nancee M.; Levine, Alan D.

doi:10.1021/bi00220a011

Cited by 41 publications

(20 citation statements)

References 33 publications

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“…Of particular interest in the light of the present structural conclusions is a comparison of the human IL-4 sequence with that of the mouse protein (DeFrance et al, 1987a;Carr et al, 1991). In the latter protein, sequence alignments indicate the absence of the 3-127 disulfide bond and its replacement with a disulfide bond linking residues 3 and 92 (human sequence numbers).…”

Section: Discussionmentioning

confidence: 82%

Secondary structure and topology of human interleukin 4 in solution

Redfield¹,

Smith²,

Boyd³

et al. 1991

Biochemistry

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Human interleukin 4 (IL-4) has been studied by 2D and 3D NMR techniques using uniformly 15N-labeled recombinant protein. Assignment of resonances for all but 3 of the 130 residues of the recombinant protein has been achieved, enabling the secondary structure of the protein to be defined. This consists of four major alpha-helical regions and one short section of double-stranded antiparallel beta-sheet. Analysis of distance and angle restraints derived from NMR experiments has enabled the overall molecular topology to be determined. This is related to that found for other four-helix proteins but has several distinctive features including cross-linking of helices by means of three disulfide bonds and a short section of beta-sheet. The structural analysis gives support to the hypothesis that many helical cytokines have a common fold and provides a basis for understanding the biological function of IL-4.

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Section: Discussionmentioning

confidence: 82%

Secondary structure and topology of human interleukin 4 in solution

Redfield¹,

Smith²,

Boyd³

et al. 1991

Biochemistry

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“…Attaching two proteins together with a new location for the junction could improve the function of the fusion protein by allowing one of the fused proteins to interact with its target with less interference or by allowing one of the proteins to fold to a more native state. In the case of the IL4 fusion proteins, determining which mechanism is responsible for the improved (25,30). Both asparagine residues are located in loops in the IL4 molecules and, therefore, appeared to us to be equivalent.…”

Section: Methodsmentioning

confidence: 99%

A circularly permuted recombinant interleukin 4 toxin with increased activity.

Kreitman

Puri

Pastan

1994

Proc. Natl. Acad. Sci. U.S.A.

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Fusion of ligands such as growth factors to other proteins often dramatically reduces the affinity of the ligand for its receptor. With recombinant DNA techniques, the attachment point between the two proteins has until now been restricted to either the amino or the carboxyl terminus of the d. However, binding may be greatly compromised if both ends are close to the site at which the ligand binds to its receptor. To construct a single-chain growth factor fusion protein with the connection at a new site on the growth factor, we constructed a DNA fria ent encoding circularly permuted interleukin 4 (H4), termed 114(38-37 In many cases, fusion of one protein to another impairs the activity of one or both proteins. This can occur if one or both proteins require one or both termini free for optimal activity. Alternatively, connection of one protein to another can impair the ability ofeither protein to properly fold. It has been reported that transforming growth factor a, IL2, IL3, IL4, IL6, GM-CSF, and insulin-like growth factor I all bind with 20-to 250-fold reduced affinity after fusion to another protein (7,(13)(14)(15)(16). Previously, the options available for improving the function of a fusion protein were limited to switching the order of the two proteins, attaching linkers in between the two proteins, or mutating one of the proteins to decrease junctional effects (13,17).We have previously reported on the properties of chimeric toxins in which IL4 was fused to recombinant forms of Pseudomonas exotoxin (PE). The fusion proteins bind to the IL4 receptor (IL4R) with only "l4% of the affinity of native IL4 (14). Several studies have indicated that the carboxyl terminus of IL4 is important for binding (18,19); therefore, it is likely that the large toxin molecule attached to the carboxyl terminus blocks the binding of 1L4 to the IL4R. We therefore developed a strategy for fusing the two proteins in which the toxin is fused to the new carboxyl terminus of circularly permuted forms of IL4. A circular permuted protein is a mutant protein in which the termini have been fused and new termini created elsewhere in the molecule. Several circularly permuted proteins have been made but none of these have been fused to other proteins (20). We reasoned that circularly permuted IL4 (CP-IL4) could be fused to a toxin so that thejunction would be in an entirely new location and allow the IL4 to bind in a more native fashion. MATERIALS AND METHODSPlasmid Construction. pRKL4EL encodes human IL4 followed by the amino acids ELKA, contains gag-ctc-gaa-gcttga (nucleotide sequences are shown by lowercase letters to avoid confusion with single-letter amino acid symbols) at the end of the coding sequence, and was derived by PCR amplification of pWDMH4 (14) and ligation into the vector pVCDT1-anti-Tac(Fv) (21). pRKL4 encodes the 129 amino acid 14 protein purified from Escherichia coli (22) and was derived from PCR amplification of pRKLAEL and ligation into the vector pVCDT1-anti-Tac(Fv). DNA encoding the toxin used for fusion to CP...

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“…Interleukin-4 (IL-4) is a protein composed of 129 amino acids, which takes the form of four interconnected α -helices additionally stabilized by three disulfide bonds [174–176]. IL-4 is a ligand whose biological activity is mediated through a receptor system dedicated to both IL-4 and IL-13 [177].…”

Section: Anti-inflammatory Cytokinesmentioning

confidence: 99%

The Role of Inflammatory and Anti-Inflammatory Cytokines in the Pathogenesis of Osteoarthritis

Wojdasiewicz

Poniatowski

Szukiewicz

2014

Mediators of Inflammation

1,238

1,132

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Osteoarthritis (OA) is the most common chronic disease of human joints. The basis of pathologic changes involves all the tissues forming the joint; already, at an early stage, it has the nature of inflammation with varying degrees of severity. An analysis of the complex relationships indicates that the processes taking place inside the joint are not merely a set that (seemingly) only includes catabolic effects. Apart from them, anti-inflammatory anabolic processes also occur continually. These phenomena are driven by various mediators, of which the key role is attributed to the interactions within the cytokine network. The most important group controlling the disease seems to be inflammatory cytokines, including IL-1β, TNFα, IL-6, IL-15, IL-17, and IL-18. The second group with antagonistic effect is formed by cytokines known as anti-inflammatory cytokines such as IL-4, IL-10, and IL-13. The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of OA with respect to inter- and intracellular signaling pathways is still under investigation. This paper summarizes the current state of knowledge. The cytokine network in OA is put in the context of cells involved in this degenerative joint disease. The possibilities for further implementation of new therapeutic strategies in OA are also pointed.

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Disulfide assignments in recombinant mouse and human interleukin 4

Cited by 41 publications

References 33 publications

Secondary structure and topology of human interleukin 4 in solution

Secondary structure and topology of human interleukin 4 in solution

A circularly permuted recombinant interleukin 4 toxin with increased activity.

The Role of Inflammatory and Anti-Inflammatory Cytokines in the Pathogenesis of Osteoarthritis

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