Mapping of two immunodominant structures on human interferon alpha 2c and their role in binding to cells

Kontsek, Peter; Borecky, adislav; Kontseková, Eva; Macikov, Ivana; Kolcunova, Alexandra; Novák, Michal; Krchňák, Viktor

doi:10.1016/0161-5890(91)90016-d

Cited by 37 publications

(9 citation statements)

References 14 publications

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“…Several reports have proposed that two conservative amino acid regions 30 -41 and 120 -145 appear to constitute the basic framework of receptor binding site (14). The site in the N-terminal region would determine the binding to high-affinity receptors, and the site in the C-terminal region would influence low-affinity binding to cells (33,34). Extensive evidence suggests that this receptor system is complex, possibly consisting of either multiple receptors or a multi-subunit receptor (22,35).…”

Section: Discussionmentioning

confidence: 99%

Human IFN-α Protein Engineering: The Amino Acid Residues at Positions 86 and 90 Are Important for Antiproliferative Activity

Bekisz

Schmeisser

et al. 2001

The Journal of Immunology

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Human IFN-α is a family of structurally related proteins that exhibit a wide range of antiproliferative activities. To understand the structural basis for these different antiproliferative activities, eight recombinant human IFN-α hybrids (HY) of α21a/α2c (HY-4, HY-5) and mutants (site-directed mutagenesis (SDM)-1, 2 and cassette mutagenesis (CM)-1, 2, 3, and 4) have been expressed, purified, and characterized. The data showed that the amino acid region 81–95 is important for antiproliferative activity. Site-directed mutagenesis and cassette mutagenesis studies showed that if serine (S) 86 and asparagine (N) 90 were replaced by tyrosine (Y), the antiproliferative activity was increased. We have also observed that if Y86 was replaced by isoleucine (I), the antiproliferative activity was comparable. However, if Y86 was replaced by aspartic acid (D), lysine (K), or alanine (A), the antiproliferative activity was substantially decreased. Our results indicate that Y and/or I at position 86 and Y at position 90 are very important in antiproliferative activity of human IFN-α. Circular dichroism spectra showed that the amino acid replacements at position 86 did not change the secondary structure. Thus the biological activity changes among those mutants do not appear to be due to conformational changes. The results also suggest that hydrophobic residue(s) at position 86 may be important for the interaction of the molecule with its receptor. The competitive binding data correlated with the antiproliferative activity. The N-terminal region of the molecule and the hydrophobic residues (including Y and I) on the C-helix region at positions 86 and/or 90 are important for binding and antiproliferative activities of human IFN-αs.

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Section: Discussionmentioning

confidence: 99%

Human IFN-α Protein Engineering: The Amino Acid Residues at Positions 86 and 90 Are Important for Antiproliferative Activity

Bekisz

Schmeisser

et al. 2001

The Journal of Immunology

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“…(16)(17)(18) (11,17,(19)(20)(21)(22) and were used as hybridoma culture supernatants containing the respective antibodies at a concentration of 5-10 mg/ml.…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Amino Acid Substitutions in Loop BC and Helix C Affect Antigenic Properties of Helix D in Hybrid IFN-α21a/α2c Molecules

Schmeisser

Kontsek

et al. 2002

Journal of Interferon & Cytokine Research

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We compared the antigenic properties of human interferon-alpha2c (IFN-alpha2c), IFN-alpha21a, hybrids IFN-alpha21a/alpha2c, and their mutants, using a panel of 27 anti-IFN-alpha1, anti-IFN-alpha2, and anti-IFN-alpha8/1/8 monoclonal antibodies (mAb). After immunoanalysis by ELISA, we found parental IFN-alpha2c and IFN-alpha21a to be antigenically distinct. Lack of reactivity of anti-IFN-alpha1 mAb with IFN-alpha21a indicated an antigenic distinction between subtypes alpha1 and alpha21a. The antigenic properties of hybrid IFNs consisting of the N-terminal portion (1-75) of IFN-alpha21a and the C-terminal portion (76-166) of IFN-alpha2c were analyzed with mAb recognizing defined regions of IFN-alpha2c, IFN-alpha1, and IFN-alpha8/1/8. We found that extending the sequence of IFN-alpha21a up to position 95 in hybrid molecule decreased the immunoreactivity of mAb specific for the antigenic structure formed by residues --112-132-- (helix D) of IFN-alpha2c. Inserting the sequence 76-81 (loop BC) of IFN-alpha2c into the sequence of 1-95 of IFN-alpha21a restored the reactivity of anti-IFN-alpha2c mAb. Some amino acid substitutions at positions 86 and 90 (helix C) of hybrid IFN-alpha21a/alpha2c also affected the immunoreactivity of C-terminal-specific mAb, which recognize helix D, but did not influence the structure of C-terminus of IFN (aa 151-165). Changes in the structure of constructs affected not only their antiproliferative activity but also their antiviral activity on human cells.

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“…Thus, unique IFN-ζ/limitin characters such as the restricted biological activities are likely to reflect differences in the receptor interactions and/or the utilization of Jak kinases after ligandreceptor binding. Experiments using monoclonal antibodies against IFN-α or IFN-β and site-directed mutagenesis revealed the functional importance of the N-terminal half of the loop AB and the loop DE together with the nearest segments of the helices D and E [50][51][52]. Another analysis using hybrid IFNs with combinations of common restriction sites suggested that the helix C interacts directly with IFNAR-1 [53].…”

Section: Studies From Ifn-ζ ζ/Limitinmentioning

confidence: 99%

IFN-? limitin: a member of type I IFN with mild lympho-myelosuppression

Oritani

Kanakura

2005

J Cellular Mol Med

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Interferon (IFN)‐ζlimitin has been considered as a novel type I IFN by the Nomenclature Committee of the International Society for Interferon and Cytokine Research. IFN‐ζlimitin shows some sequence homology with IFN‐α and IFN‐β, has a globular structure with five α‐helices and four loops, and recognizes IFN‐α/β receptor. Although IFN‐ζlimitin displays antiviral, immunomodulatory, and antitumor effects, it has much less lymphomyelosuppressive activities than IFN‐α. Treatment of cells with type I IFNs induces and/or activates a number of molecules, which regulate cell cycle and apoptosis. It is noteworthy that IFN‐ζlimitin activates the Tyk2‐Daxx and Tyk2‐Crk pathways weaker than IFN‐α. Because experiments using antisense oligonucleotides have revealed their essential role in type I IFN‐related suppression of lympho‐hematopoiesis, little ability of IFN‐ζlimitin to activate the Tyk2‐dependent signaling pathway may explain its uniquely narrow range of biological activities. Further analysis of structure‐function relationship of type I IFNs will establish an engineered cytokine with useful features of IFN‐ζlimitin.

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Mapping of two immunodominant structures on human interferon alpha 2c and their role in binding to cells

Cited by 37 publications

References 14 publications

Human IFN-α Protein Engineering: The Amino Acid Residues at Positions 86 and 90 Are Important for Antiproliferative Activity

Human IFN-α Protein Engineering: The Amino Acid Residues at Positions 86 and 90 Are Important for Antiproliferative Activity

Amino Acid Substitutions in Loop BC and Helix C Affect Antigenic Properties of Helix D in Hybrid IFN-α21a/α2c Molecules

IFN-? limitin: a member of type I IFN with mild lympho-myelosuppression

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