Conformational effects of amino acid substitutions at positions 10, 12, and 13 in the P21 protein.
Substitutions of amino acids for Gly 12 or Gly 13 in the ras oncogene-encoded P21 proteins have been demonstrated to produce unique structural changes in these proteins that correlate with their ability to produce cell transformation. For example, the P21 proteins with Arg 12 or Val 13 are both known to be actively transforming. Recent site-specific mutagenesis experiments on the transforming Arg 12 protein have found that the substitution of Val for Gly 10 has no effect on transforming activity whereas the substitution of Val for Gly 13 led to a loss of transforming activity. In this study, we examine the structural effects of these substitutions on the amino terminal hydrophobic decapeptide (Leu 6-Gly 15) of P21 using conformational energy analysis. The results show that the transforming proteins with Gly 10 and Arg 12 or Val 10 and Arg 12 can both adopt the putative malignancy-causing conformation, whereas, for the nontransforming protein with Arg 12 and Val 13, this conformation is energetically disallowed. These results further support the theory that due to structural changes the transforming P21 proteins are unable to bind to some regulatory cellular element which may be the recently identified binding protein responsible for the induction of increased GTPase activity in normal P21 compared with transforming mutants.
Brandt-Rauf
P W
PW
Division of Environmental Sciences, Columbia University, New York, New York 10032.
Pincus
M R
MR
Carty
R P
RP
Lubowsky
J
J
Avitable
M
M
Kung
H F
HF
Maizel
J
J
D000596:Amino Acids / Q000032:analysis*; D001665:Binding Sites; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002942:Circular Dichroism; D004735:Energy Transfer; D020558:GTP Phosphohydrolases / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000032:analysis; D011487:Protein Conformation; D011518:Proto-Oncogene Proteins / Q000032:analysis* / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D012984:Software; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014961:X-Ray Diffraction
D000596:Amino Acids; D010455:Peptides; D011518:Proto-Oncogene Proteins; D020558:GTP Phosphohydrolases; D016283:Proto-Oncogene Proteins p21(ras)