Molecular determinants for the complex formation between the retinoblastoma protein and LXCXE sequences.
The retinoblastoma tumor suppressor protein (pRb) is a key negative regulator of cell proliferation that is frequently disregulated in human cancer. Many viral oncoproteins (for example, HPV E7 and E1A) are known to bind to the pRb pocket domain via a LXCXE binding motif. There are also some 20 cellular proteins that contain a LXCXE motif and have been reported to associate with the pocket domain of pRb. Using NMR spectroscopy and isothermal calorimetry titration, we show that LXCXE peptides of viral oncoproteins bind strongly to the pocket domain of pRb. Additionally, we show that LXCXE-like peptides of HDAC1 bind to the same site on pRb with a weak (micromolar) and transient association. Systematic substitution of residues other than conserved Leu, Cys, and Glu show that the residues flanking the LXCXE are important for the binding, whereas positively charged amino acids in the XLXCXEXXX sequence significantly weaken the interaction.
Singh
Mahavir
M
Max Planck Institute for Biochemistry, Martinsried, Germany.
Krajewski
Marcin
M
Mikolajka
Aleksandra
A
Holak
Tad A
TA
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D008958:Models, Molecular; D009856:Oncogene Proteins, Viral / Q000737:chemistry* / Q000378:metabolism*; D050725:Papillomavirus E7 Proteins / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016160:Retinoblastoma Protein / Q000737:chemistry* / Q000378:metabolism*
D000952:Antigens, Polyomavirus Transforming; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D010446:Peptide Fragments; D016160:Retinoblastoma Protein