The structure of an antigenic determinant in a protein

Wilson, Ian A.; Niman, Henry L.; Houghten, Richard A.; Cherenson, Andrew R.; Connolly, Michael L.; Lerner, Richard A.

doi:10.1016/0092-8674(84)90412-4

Cited by 817 publications

(537 citation statements)

References 31 publications

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“…A nine amino acid epitope tag from the in¯uenza virus hemagglutin protein (HA) (Wilson et al, 1984) was added to the N-terminus of the full length long and short forms of cbl-3 by PCR and the cDNAs were cloned into pCEFL, a mammalian expression vector with the elongation factor promoter and a neomycin selectable marker (provided by Dr Silvio Gutkind). The construct was sequenced to verify that there were no mutations introduced.…”

Section: Cell Culture and Transfectionsmentioning

confidence: 99%

cbl-3: a new mammalian cbl family protein

et al. 1999

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We have cloned a new human gene, cbl-3, which encodes a protein with marked homology to the cbl family of proteins. The predicted protein encoded by this gene retains the conserved phosphotyrosine binding domain (PTB) in the N-terminal and the zinc ®nger but is signi®cantly shorter (MW 52.5 kDa) than the other mammalian cbl proteins. The protein lacks the extensive proline rich domain and leucine zipper seen in c-cbl and cbl-b and structurally most resembles the C. elegans and Drosophila cbl proteins. The gene is ubiquitously expressed with highest expression in the aerodigestive tract, prostate, adrenal gland, and salivary gland. The protein is phosphorylated and recruited to the EGFR upon EGF stimulation and inhibits EGF stimulated MAP kinase activation. In comparison to the other mammalian cbl proteins (e.g. cbl-b), cbl-3 interacts with a restricted range of proteins containing Src Homology 3 regions. An alternatively spliced form of the cbl-3 protein was also identi®ed which deletes a critical region of the PTB domain and which does not interact with the EGFR nor inhibit EGF stimulated MAP kinase activation. These data demonstrate that cbl-3, a novel mammalian cbl protein, is a regulator of EGFR mediated signal transduction.

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Section: Cell Culture and Transfectionsmentioning

confidence: 99%

cbl-3: a new mammalian cbl family protein

et al. 1999

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“…A nine amino acid epitope tag from the in¯uenza virus hemagglutin protein (HA) (Wilson et al, 1984) was added to the C-terminal of the full length human cbl-b open reading frame (Keane et al, 1995) by PCR and the cDNA was cloned into pCEFL, a mammalian expression vector with the elongation factor promoter and a neomycin selectable marker (provided by Dr Silvio Gutkind). The construct was sequenced to verify that there were no mutations introduced.…”

Section: Plasmidsmentioning

confidence: 99%

cbl-b inhibits epidermal growth factor receptor signaling

et al. 1999

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The role of cbl-b in signaling by the epidermal growth factor receptor (EGFR) was studied and compared with c-cbl. We demonstrate in vivo, that cbl-b, like c-cbl, is phosphorylated and recruited to the EGFR upon EGF stimulation and both cbl proteins can bind to the Grb2 adaptor protein. To investigate the functional role of cbl proteins in EGFR signaling, we transfected cbl-b or c-cbl into 32D cells overexpressing the EGFR (32D/EGFR). This cell line is absolutely dependent on exogenous IL-3 or EGF for sustained growth. 32D/EGFR cells overexpressing cbl-b showed markedly inhibited growth in EGF compared to c-cbl transfectants and vector controls. This growth inhibition by cbl-b was the result of a dramatic increase in the number of cells undergoing apoptosis. Consistent with this ®nding, cbl-b overexpression markedly decreased the amplitude and duration of AKT activation upon EGF stimulation compared to either vector controls or c-cbl overexpressing cells. In addition, the duration of EGF mediated MAP kinase and Jun kinase activation in cells overexpressing cbl-b is shortened. These data demonstrate that cbl-b inhibits EGF-induced cell growth and that cbl-b and c-cbl have distinct roles in EGF mediated signaling.

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“…Anti-Ret antibodies were used at 1 mg/ml for immunoprecipitation and at 0.1 mg/ml for Western blotting. Mouse monoclonal antibodies anti-HA epitope (clone 12CA5) (Wilson et al, 1984) were purchased from Boehringer Mannheim, Mannheim, Germany. Rabbit polyclonal anti-JNK antiserum was purchased from Santa Cruz Biotechnology, CA.…”

Section: Expression Plasmids and Antibodiesmentioning

confidence: 99%

Signalling of the Ret receptor tyrosine kinase through the c-Jun NH2-terminal protein kinases (JNKs): evidence for a divergence of the ERKs and JNKs pathways induced by Ret

et al. 1998

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The RET proto-oncogene encodes a functional receptor tyrosine kinase (Ret) for the Glial cell line Derived Neurotrophic Factor (GDNF). RET is involved in several neoplastic and non-neoplastic human diseases. Oncogenic activation of RET is detected in human papillary thyroid tumours and in multiple endocrine neoplasia type 2 syndromes. Inactivating mutations of RET have been associated to the congenital megacolon, i.e. Hirschprung's disease. In order to identify pathways that are relevant for Ret signalling to the nucleus, we have investigated its ability to induce the c-Jun NH 2 -terminal protein kinases (JNK). Here we show that triggering the endogenous Ret, expressed in PC12 cells, induces JNK activity; moreover, Ret is able to activate JNK either when transiently transfected in COS-1 cells or when stably expressed in NIH3T3 ®broblasts or in PC Cl 3 epithelial thyroid cells. JNK activation is dependent on the Ret kinase function, as a kinase-de®cient RET mutant, associated with Hirschsprung's disease, fails to activate JNK. The pathway leading to the activation of JNK by RET is clearly divergent from that leading to the activation of ERK: substitution of the tyrosine 1062 of Ret, the Shc binding site, for phenylalanine abrogates ERK but not JNK activation. Experiments conducted with dominant negative mutants or with negative regulators demonstrate that JNK activation by Ret is mediated by Rho/Rac related small GTPases and, particularly, by Cdc42.

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The structure of an antigenic determinant in a protein

Cited by 817 publications

References 31 publications

cbl-3: a new mammalian cbl family protein

cbl-3: a new mammalian cbl family protein

cbl-b inhibits epidermal growth factor receptor signaling

Signalling of the Ret receptor tyrosine kinase through the c-Jun NH2-terminal protein kinases (JNKs): evidence for a divergence of the ERKs and JNKs pathways induced by Ret

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