Identification of the sites of interaction between c-Raf-1 and Ras-GTP.

@article{citeulike:2801249, abstract = {Specific sites of protein-protein interaction were identified in the 51-149 region of c-Raf-1 using contact epitope scanning and site-directed mutagenesis. Nineteen overlapping peptides based upon the primary sequence of the Ras binding domain of c-Raf-1 were tested for the ability to competitively inhibit complex formation between Ras-GTP and the c-Raf-1 N-terminus. A peptide containing c-Raf-1 residues 91-105 as well as five overlapping peptides covering a region extending from residues 118 to 143 interfered with Ras association, defining these sites as potential contact surfaces with Ras. Alanine scanning mutagenesis was used as a second probe for sites of Ras interaction with the c-Raf-1 N-terminus. Raf residues 64-67 and 80-103 were demonstrated as important for association with Ras-GTP with residues 66, 67, 84, 87, 89 and 91 identified as the most critical individual points of contact with the Ras protein. Alanine substitution of residues between 118-143 suggested only one potentially weak site of interaction defined by residues 120-125. The combined results of both peptide and mutagenic analyses suggest that the primary site of c-Raf-1 interaction with Ras maps to Raf residues 80-103, with secondary interactions occurring with residues 66 and 67 and possibly 120-125. Contact epitope scanning of the Ras effector region found maximum inhibition of Ras/Raf association with a peptide corresponding to Ras amino acids 37-51. A model is proposed for the GTP-dependent association of Ras and Raf.}, address = {Department of Medicine, Indiana University, Indianapolis 46202, USA.}, author = {Barnard, D. and Diaz, B. and Hettich, L. and Chuang, E. and Zhang, X. F. and Avruch, J. and Marshall, M. }, citeulike-article-id = {2801249}, issn = {0950-9232}, journal = {Oncogene}, keywords = {ras, usa}, month = {April}, number = {7}, pages = {1283--1290}, posted-at = {2008-05-15 11:25:00}, priority = {2}, title = {Identification of the sites of interaction between c-Raf-1 and Ras-GTP.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/7731678}, volume = {10}, year = {1995} }

TY - JOUR ID - citeulike:2801249 L3 - citeulike-article-id:2801249 TI - Identification of the sites of interaction between c-Raf-1 and Ras-GTP. JF - Oncogene VL - 10 IS - 7 SP - 1283 EP - 1290 AD - Department of Medicine, Indiana University, Indianapolis 46202, USA. SN - 0950-9232 N2 - Specific sites of protein-protein interaction were identified in the 51-149 region of c-Raf-1 using contact epitope scanning and site-directed mutagenesis. Nineteen overlapping peptides based upon the primary sequence of the Ras binding domain of c-Raf-1 were tested for the ability to competitively inhibit complex formation between Ras-GTP and the c-Raf-1 N-terminus. A peptide containing c-Raf-1 residues 91-105 as well as five overlapping peptides covering a region extending from residues 118 to 143 interfered with Ras association, defining these sites as potential contact surfaces with Ras. Alanine scanning mutagenesis was used as a second probe for sites of Ras interaction with the c-Raf-1 N-terminus. Raf residues 64-67 and 80-103 were demonstrated as important for association with Ras-GTP with residues 66, 67, 84, 87, 89 and 91 identified as the most critical individual points of contact with the Ras protein. Alanine substitution of residues between 118-143 suggested only one potentially weak site of interaction defined by residues 120-125. The combined results of both peptide and mutagenic analyses suggest that the primary site of c-Raf-1 interaction with Ras maps to Raf residues 80-103, with secondary interactions occurring with residues 66 and 67 and possibly 120-125. Contact epitope scanning of the Ras effector region found maximum inhibition of Ras/Raf association with a peptide corresponding to Ras amino acids 37-51. A model is proposed for the GTP-dependent association of Ras and Raf. KW - ras KW - usa AU - Barnard, D AU - Diaz, B AU - Hettich, L AU - Chuang, E AU - Zhang, XF AU - Avruch, J AU - Marshall, M PY - 1995/04/06/ UR - http://view.ncbi.nlm.nih.gov/pubmed/7731678 ER -