Posttranslational modification of human T-cell growth factor

Robb, Richard J.; Kutny, Rusty; Panico, Maria; Morris, Howard R.; DeGrado, William F.; Chowdhry, Vinay

doi:10.1016/s0006-291x(83)80248-4

Cited by 41 publications

(8 citation statements)

References 17 publications

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“…An earlier study suggested that glycosylation of Thr-3 was involved in recognition of IL2 by the 1H11-1A5 antibody (9). Indeed, the Aff-IL2 NH2-terminal octapeptide with the GaINAc modification was as effective on a molar basis as intact IL2 in competing with the binding of radiolabeled IL2 to antibody-coupled Sepharose beads.…”

Section: Results --mentioning

confidence: 99%

Amino acid sequence and post-translational modification of human interleukin 2.

Robb¹,

Kutny²,

Panico³

et al. 1984

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

106

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Human interleukin 2 was separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. For the most part, this heterogeneity was attributed to variations in glycosylation of the threonine residue in position 3 of the polypeptide chain. The various molecular forms of interleukin 2 had nearly identical specific activities in the in vitro proliferation assay, indicating that the glycosylation had no significant effect on this response. The entire primary sequence of interleukin 2, including the location of the intramolecular disulfide bridge, was determined by a combination of peptide mapping and protein sequencing. This information should aid in the determination of the active site(s) of the molecule.

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Section: Results --mentioning

confidence: 99%

Amino acid sequence and post-translational modification of human interleukin 2.

Robb¹,

Kutny²,

Panico³

et al. 1984

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

106

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“…Examples of fucose present as a branched carbohydrate structure and linked to N-acetylglucosamine have been reported (7). However, we know of no precedent for a direct fucosylation of peptide in a distinct biologically active protein, although attachment of the monosaccharide N-acetyl-D-galactosamine to threonine at position 3 of human T-cell growth factor has been reported (23). A direct covalent bond between fucose and the amino acid threonine has been reported (24), but such linkage has not been described for any distinct protein, nor has the site of modification been defined (8,25).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a posttranslational fucosylation in the growth factor domain of urinary plasminogen activator.

Buko

Kentzer

Petros

et al. 1991

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

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A posttranslational modification site in natural and recombinant urinary-type plasminogen activators (urokinases; EC 3.4.21.31) has been localized to Thr-18, in the growth factor domain of the molecule. This is the region of urinary plasminogen activator responsible for its specific receptor binding. An unusual carbohydrate-protein linkage, a single monosaccharide, fucose, covalently attached directly to threonine in the peptide, is described here. The glycan moiety and the site of modification have been identified with mass spectrometry and confirmed by carbohydrate composition analysis, Edman degradation, and one-and two-dimensional NMR studies. This type of modification is normally not detected without mass spectrometry because the fucose-threonine bond is hydrolyzed under standard acidic conditions of the amino acid analysis and Edman sequencing. This modification may be widely found in other proteins.

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“…Highly purified recombinant IL2 (>95%) used in our study was kindly provided by (Robb et al, 1983) was kindly supplied by Dr. R.J. Robb (du Pont de Nemours, Glenolden, PA). OKT3 was purchased from Ortho (Roissy, France).…”

Section: Lak Activation and Cytotoxicity Assaymentioning

confidence: 99%

“…In an attempt to evaluate the role of the p55 and the p75 peptides in the increase of TNF binding sites on LGL and the subsequent LAK promotion we performed blocking studies using anti-Tac antibodies known to block the interaction of IL2 with the high-affinity receptors (Robb et aE., 1984;Trinchieri et aE., 1984) and 1 HI 1 MAb that specifically inhibits binding of IL2 to the p75 component (Robb et al, 1983). To this end, LGL were cultured with IL2 (0.25 ng/ml) and TNF (250 ng/ml) in the presence of saturating amounts of anti-Tac antibody (25 pg/ml) or l H l l MAb (15 pglml).…”

Section: Involvement Of the P55 And The P75 Chains In The Triggering mentioning

confidence: 99%

Functional interactions of IL2 and TNF in the differentiation of LGL into lak effectors

Blay

Bertoglio

Fradelizi

et al. 1989

Intl Journal of Cancer

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We have previously reported on the synergistic effect of IL2 and TNF on the differentiation of LGL into LAK effectors. In the present study, we have further investigated the molecular basis of this synergistic mechanism and examined the role of TNF in the induction of LAK activity. We show that the generation of optimal LAK activity by low doses of IL-2 in the presence of TNF involves the induction of high-affinity IL2 receptors on LGL and occurs without promoting significant proliferation, suggesting a functional activation rather than a proliferative expansion of LAK precursors. Using blocking studies with anti-Tac and with an anti-IL2 (IHII), which specifically inhibits the binding of IL2 to the p75 IL2 receptor component, we also show that both the p55 and the p75 are involved in the increase in TNF binding sites on LGL and the subsequent acquisition of LAK activity. We also demonstrate that the failure of low doses of IL2 to induce LAK activity is related to their incapacity to induce TNF production. Moreover, when specific antibodies against TNF were added to the culture, the differentiation of LGL into LAK effectors by optimal concentrations of IL2 in our system (2.5-5.0 ng/ml) was partially inhibited. This suggests that TNF may be a physiologic mediator in the sequential activation stages of LGL into LAK effectors.

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Posttranslational modification of human T-cell growth factor

Cited by 41 publications

References 17 publications

Amino acid sequence and post-translational modification of human interleukin 2.

Amino acid sequence and post-translational modification of human interleukin 2.

Characterization of a posttranslational fucosylation in the growth factor domain of urinary plasminogen activator.

Functional interactions of IL2 and TNF in the differentiation of LGL into lak effectors

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