Amino acid sequence of a human leukocyte interferon.

Levy, Warren P.; Rubinstein, Menachem; Shively, John E.; Valle, U Del; Lai, Chun‐Yen; Moschera, John; Brink, Larry; Gerber, Louise; Stein, Stanley J.; Pestka, Sidney

doi:10.1073/pnas.78.10.6186

Cited by 102 publications

(23 citation statements)

References 32 publications

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“…This suggests that the MPF precursor would fold and then be successively processed with furin-like and trypsin-like proteases. A similar processing has been found in leukocyte-derived natural IFN-a (27) and IFN-g (28,29).…”

Section: Discussionmentioning

confidence: 76%

Mesothelin Variant 1 Is Released from Tumor Cells as a Diagnostic Marker

Hellström

Raycraft

Kanan

et al. 2006

Cancer Epidemiology, Biomarkers &Amp; Prevention

102

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The mesothelin family comprises (at least) three variants and includes the precursor for megakaryocyte potentiating factor (MPF). Assaying soluble mesothelin-related protein (SMRP) molecules in serum and other body fluids from patients with certain cancers can provide diagnostically useful information. We have constructed fusion proteins of mesothelin variants 1, 2, and 3, made monoclonal antibodies, and investigated the binding specificity of these and three previously generated monoclonal antibodies to each of the three mesothelin variants. According to flow cytometry, the molecule that is most frequently expressed at the surface of cells from ovarian carcinomas and certain other tumors is mesothelin variant 1. Similarly, SMRP released into ascites from a patient with ovarian carcinoma was shown to have a molecular weight of f40 kDa and, according to sequencing, to be variant 1. A published sandwich ELISA was shown to detect variants 1 and 3 and to be much more sensitive than a newly constructed ELISA, which detects only variant 3, the former being positive in 28 of 41 (68%) sera from patients with ovarian cancer as compared with 6 of 41 sera (15%). A standard curve was constructed to measure SMRP with a limit of detection of 200 pg/mL to facilitate future quantitative studies. (Cancer Epidemiol Biomarkers Prev 2006;15(5):1014 -20)

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Section: Discussionmentioning

confidence: 76%

Mesothelin Variant 1 Is Released from Tumor Cells as a Diagnostic Marker

Hellström

Raycraft

Kanan

et al. 2006

Cancer Epidemiology, Biomarkers &Amp; Prevention

102

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“…Human leukocyte interferon (IFN-a) is a family of individual species differing in amino acid sequences (1)(2)(3). Several recombinant IFN-a molecules and hybrid IFNs have been expressed in Escherichia coli, purified to homogeneity, and characterized (1,(4)(5)(6).…”

mentioning

confidence: 99%

A species of human alpha interferon that lacks the ability to boost human natural killer activity.

Ortaldo¹,

Herberman²,

Harvey³

et al. 1984

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

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100

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Most species of recombinant leukocyte interferons (IFN-aA, -aB, -aC, -aD, -aF, -61, and -aK) were capable of boosting human natural killer (NK) activity after a 2-hr treatment of cells at a concentration of 1-80 units/ml. In contrast, recombinant human IFN-azJ was found to be incapable of augmenting NK activity after exposure of cells for 2 hr to concentrations as high as 10,000 units/ml. This inability of IFN-aJ to boost NK activity was not complete because, after exposure of cells to a high concentration of IFN-aJ (10,000 units/ml) for 18 hr, boosting of cytolysis was observed. IFNaJ appeared to interact with receptors for IFN on NK cells since it was found to interfere with the boosting of NK activity by other species of IFN-a. In contrast to its deficient ability to augment NK activity, IFN-ErJ has potent antiviral and antiproliferative activities. Such extensive dissociation of these biological activities has not been observed previously with any other natural or recombinant IFN species. Thus, this IFN species may be useful for evaluating the relative importance of various biological activities on the therapeutic effects of IFN, for understanding structure-function relationships, and for determining the biochemical pathways related to the various biological effects of IFN.Human leukocyte interferon (IFN-a) is a family of individual species differing in amino acid sequences (1-3). Several recombinant IFN-a molecules and hybrid IFNs have been expressed in Escherichia coli, purified to homogeneity, and characterized (1, 4-6). These recombinant and hybrid IFNs as well as natural IFNs vary considerably in their patterns of antiviral activity on human, bovine, mouse, feline, and rat cells. IFNs also have been shown to modify a wide variety of biological responses, and previous studies (1, 7-11) have indicated substantial quantitative differences among the natural and recombinant IFN species in their relative potency in exerting antiviral, antiproliferative, and immunomodulating activities. Previous reports (9, 12--16) have demonstrated that natural killer (NK) cell activity can be significantly augmented by natural and recombinant IFN-a species. However, each of the species previously tested exhibited an ability to mediate all of these effects. In this report we describe the dissociation of the antiviral activities of IFN-aJ from its ability to stimulate NK cells.

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“…Aliquots were then reduced with electrophoresis solubilizing buffer containing 3% 2-mercaptoethanol and applied to 16% polyacrylamide gels containing 0.1% NaDodSO4. ' -- (3,6,19,20). Studies with a monoclonal antibody, which recognizes the last 16 carboxyl-terminal amino acid residues of HuIFN-aA, have confirmed that the carboxyl-terminal region of the HuIFNaA molecule is not involved in interaction with the receptor, for this antibody is capable of binding to HuIFN-aA bound to its receptor (21,22).…”

Section: Resultsmentioning

confidence: 89%

Biologic activity in a fragment of recombinant human interferon alpha.

Ackerman¹,

Nedden²,

Heintzelman³

et al. 1984

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

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To attempt to locate functionally important regions of the interferon (IFN) molecule, recombinant human IFN-a2 was subjected to proteolytic digestion. The bacterial proteinase thermolysin produced two major complementary fragments, and . After reduction with 2-mercaptoethanol and separation of the two major fragments on NaDodSO4/polyacrylamide gel electrophoresis, antiviral activity persisted in the larger, Mr 12,000, fragment consisting of the amino-terminal 110 amino acids.The availability of large amounts of recombinant DNA-derived interferon (IFN) has fostered investigation of IFN's structure-function relationships; however, the specific structural features required for biologic activity remain largely unknown. Despite disul'ide bond assignment (1), secondary structure measurements (2), and investigation of novel recombinant IFNs, (3, 4), little information is available as to which areas of the protein may be functionally important. Modification of the structure of IFNs by using a variety of enzymes and chemicals has yielded varied results. Incubation of IFNs with glycosidic enzymes in general does not appear to alter the antiviral activity (5). Cleavage of IFNs with endopeptidases such as trypsin or with chemicals such as cyanogen bromide has thus far been unsuccessful in generating active fragments (6, 7). In contrast, some active species were obtained after treatment with exopeptidases (8), pepsin (9), and periodate (10); however, reductions in size were modest with the active species of Mr 15,000 or larger. To attempt to identify small functional portions of the IFN molecule we have studied proteolytic fragments of a recombinant human IFN, HuIFN-a2, produced by the bacterial proteinase thermolysin. We report here that proteolysis of this IFN with thermolysin generates a biologically active fragment originating from the amino-terminal end of the molecule. MATERIALS AND METHODSThermolysin was obtained from Calbiochem. HuIFN-a2 (1.5x 10' units/mg of protein) was the gift of Schering. HuIFNaA (2.0 x 108 units/mg of protein) was the gift of HoffmannLa Roche. 125I-labeled HuIFN-aA (1251-HuIFN-aA) was prepared as described (11).Preparative NaDodSO4/polyacrylamide gel electrophoresis (NaDodSO4/PAGE) was performed according to the method of Laemmli (12). The separating gels were 16% acrylamide/N,N'-methylene bisacrylamide, 0.75 mm in thickness and 12 cm in length. Electrophoresis was performed at 15 mA per slab gel. The gels were stained and destained as described (13). The stained gel bands were sliced from the gel and eluted as reported (13). The samples were then assayed for biological activity or dialyzed and lyophilized for amino acid analysis and amino acid sequence determination as described (14). Continuous elution of IFN fragments was done by the method of Hunkapiller (unpublished data).IFN antiviral assays were performed by using Madin-Darby bovine kidney (MDBK) cells or human foreskin fibroblasts with vesicular stomatitis virus as reported (15). Competitive receptor binding assays using 125I-H...

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Amino acid sequence of a human leukocyte interferon.

Cited by 102 publications

References 32 publications

Mesothelin Variant 1 Is Released from Tumor Cells as a Diagnostic Marker

Mesothelin Variant 1 Is Released from Tumor Cells as a Diagnostic Marker

A species of human alpha interferon that lacks the ability to boost human natural killer activity.

Biologic activity in a fragment of recombinant human interferon alpha.

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