Antibody of predetermined specificity to a carboxy-terminal region of H-ras gene products inhibits their guanine nucleotide-binding function.

Srivastava, Shiv; Lacal, Juan Carlos; Reynolds, Steven H.; Aaronson, S A

doi:10.1128/mcb.5.11.3316

Cited by 12 publications

(8 citation statements)

References 31 publications

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“…The functional connection between this lipid modification and Ras function was made by Douglas Lowy’s group in 1984, which showed that lipid binding and membrane association were actually required for the transforming activity of the viral H-Ras oncoprotein 49,50 . working with cellular H-Ras, Stuart Aaronson’s group proceeded to demonstrate that this C-terminal processing and membrane recruitment of Ras is a prerequisite to its biochemical activation 51 .…”

Section: Post-translational Modificationsmentioning

confidence: 99%

Ras oncogenes: split personalities

Karnoub

Weinberg

2008

Nat Rev Mol Cell Biol

1,318

1,238

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Extensive research on the Ras proteins and their functions in cell physiology over the past 30 years has led to numerous insights that have revealed the involvement of Ras not only in tumorigenesis but also in many developmental disorders. Despite great strides in our understanding of the molecular and cellular mechanisms of action of the Ras proteins, the expanding roster of their downstream effectors and the complexity of the signalling cascades that they regulate indicate that much remains to be learnt.

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Section: Post-translational Modificationsmentioning

confidence: 99%

Ras oncogenes: split personalities

Karnoub

Weinberg

2008

Nat Rev Mol Cell Biol

1,318

1,238

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“…However, these studies indicate the biochemical reactivity of the polyclonal sera with their respective proteins was only fair at best and in the case of one polyclonal sera it was reported not to react with the p21 at all. Additional studies have reported antisera to the amino acid sequence 161-176 of the Ha-ras p21 (13) while another report describes antisera raised to a synthetic peptide representing amino acids 171-186 of the Ki-ras p21 (14).…”

Section: Discussionmentioning

confidence: 97%

“…To date, there have been several reports describing polyclonal sera to the individual Ha, Ki and N-ras p21 (12)(13)(14)(15). However, the limited availability of these reagents and the inconsistencies that exist between the different polyclonal sera have restricted investigations of the Ha, Ki and N-ras p21.…”

Section: Introductionmentioning

confidence: 97%

Production and Characterization of Monoclonal Antibodies to Ha-ras and N-ras p21

Conlon²,

LaVecchio³

et al. 1991

Hybridoma

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The mammalian ras family consists of the Ha, Ki and N-ras genes that encode a series of 21,000 dalton proteins (p21). The three ras proteins participate in normal cell physiology and have been implicated in cellular transformation by either overexpression of the normal p21 or by mutation at positions 12, 13, or 61. To help understand the biological roles of the different ras proteins, we have generated monoclonal antibodies (Mabs) to the Ha-ras and N-ras p21. Mab Ha-770, raised to a Ha-specific synthetic peptide, reacts with Ha-ras recombinant p21 (r-p21) as well as cellular Ha-ras p21 by immunoprecipitation. Western blot and sandwich ELISA assays. Mab N-838, raised to an N-ras specific synthetic peptide, reacts with the N-ras recombinant p21 by immunoprecipitation, Western blot and sandwich ELISA assays. Mabs to the Ha-ras and N-ras p21 should be valuable reagents in assessing the individual roles of ras proteins in normal and neoplastic cells.

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“…As yet, there has been only indirect evidence concerning the biologic importance of GTP binding in p21 function as well as the localization of GTP-binding site(s) within the p21 molecule. Such studies have relied for the most part on antibodies which inhibit biochemical activities of purified bacterially expressed ras proteins . Srivastava et al, 1985 or affect the phenotype of ras proteins expressing cells upon microinjection .…”

Section: Introductionmentioning

confidence: 99%

Deletion mutants of Harvey ras p21 protein reveal the absolute requirement of at least two distant regions for GTP-binding and transforming activities.

Lacal¹,

Anderson²,

Aaronson³

1986

The EMBO Journal

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Deletions of small sequences from the viral Harvey ras gene have been generated, and resulting ras p21 mutants have been expressed in Escherichia coli. Purification of each deleted protein allowed the in vitro characterization of GTP‐binding, GTPase and autokinase activity of the proteins. Microinjection of the highly purified proteins into quiescent NIH/3T3 cells, as well as transfection experiments utilizing a long terminal repeat (LTR)‐containing vector, were utilized to analyze the biological activity of the deleted proteins. Two small regions located at 6‐23 and 152‐165 residues are shown to be absolutely required for in vitro and in vivo activities of the ras product. By contrast, the variable region comprising amino acids 165‐184 was shown not to be necessary for either in vitro or in vivo activities. Thus, we demonstrate that: (i) amino acid sequences at positions 5‐23 and 152‐165 of ras p21 protein are probably directly involved in the GTP‐binding activity; (ii) GTP‐binding is required for the transforming activity of ras p21 and by extension for the normal function of the proto‐oncogene product; and (iii) the variable region at the C‐terminal end of the ras p21 molecule from amino acids 165 to 184 is not required for transformation.

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Antibody of predetermined specificity to a carboxy-terminal region of H-ras gene products inhibits their guanine nucleotide-binding function.

Cited by 12 publications

References 31 publications

Ras oncogenes: split personalities

Ras oncogenes: split personalities

Production and Characterization of Monoclonal Antibodies to Ha-ras and N-ras p21

Deletion mutants of Harvey ras p21 protein reveal the absolute requirement of at least two distant regions for GTP-binding and transforming activities.

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