Ribonuclease inhibitor
| Leucine Rich Repeat | |||||||||
|---|---|---|---|---|---|---|---|---|---|
 Top view of porcine ribonuclease inhibitor, showing its horseshoe shape.[1] The outer layer is composed of α-helices and the inner layer of parallel β-strands. The inner and outer diameters are roughly 2.1 nm and 6.7 nm, respectively. | |||||||||
| Identifiers | |||||||||
| Symbol | LRR_1 | ||||||||
| Pfam | PF00560 | ||||||||
| Pfam clan | CL0022 | ||||||||
| InterPro | IPR003590 | ||||||||
| SMART | SM00368 | ||||||||
| SCOP | 1bnh | ||||||||
| SUPERFAMILY | 1bnh | ||||||||
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Ribonuclease inhibitor (RI) is a large (~450 residues, ~49 kDa), acidic (pI ~4.7), leucine-rich repeat protein that forms extremely tight complexes with certain ribonucleases. It is a major cellular protein, comprising ~0.1% of all cellular protein by weight, and appears to play an important role in regulating the lifetime of RNA.[2]
RI has a surprisingly high cysteine content (~6.5%, cf. 1.7% in typical proteins) and is sensitive to oxidation. RI is also rich in leucine (21.5%, compared to 9% in typical proteins) and commensurately lower in other hydrophobic residues, esp. valine, isoleucine, methionine, tyrosine, and phenylalanine.
Structure
RI is the classic leucine-rich repeat protein, consisting of alternating α-helices and β-strands along its backbone. These secondary structure elements wrap around in a curved, right-handed solenoid that resembles a horseshoe. The parallel β-strands and α-helices form the inner and outer wall of the horseshoe, respectively. The structure appears to be stabilized by buried asparagines at the base of each turn, as it passes from α-helix to β-strand. The αβ repeats alternate between 28 and 29 residues in length, effectively forming a 57-residue unit that corresponds to its genetic structure (each exon codes for a 57-residue unit).
Binding to ribonucleases
The affinity of RI for ribonucleases is perhaps the highest for any protein-protein interaction; the dissociation constant of the RI-RNase A complex is roughly 20 fM under physiological conditions while that for the RI-angiogenin complex is even smaller (<1 fM). Remarkably, RI is able to bind a wide variety of RNases, despite having low sequence identity. Structural studies indicate that RNases bind like a "cork in the bottle", associating especially with the C-terminal end of RI; the interaction is largely electrostatic but also buries a lot of surface area (>25 nm2). Efforts to mutate RNases to lower their affinity for RI while maintaining their enzymatic activity have had limited success. However, mammalian RI seems unable to bind a few amphibian ribonucleases[citation needed], such as ranpirnase (also known as Onconase).
RI's affinity for ribonucleases is important, since ribonucleases have cytotoxic and cytostatic effects (especially against cancer cells), and are under investigation as potential cancer therapeutics. Successful evasion of the ubiquitous RI would be essential for the success of a ribonuclease drug, (since it would be ineffective bound to RI). The frog protein Onconase is under investigation for treatment of skin cancers; unfortunately, the antigenicity of amphibian proteins makes them unsuitable for treating internal human cancers. Modifications of human ribonucleases that evade RI but retain their enzymatic activity have also been studied.
References
- ↑ 1.0 1.1 PDB 2BNH; Kobe B, Deisenhofer J (1993). "Crystal structure of porcine ribonuclease inhibitor, a protein with leucine-rich repeats". Nature 366 (6457): 751–6. doi:10.1038/366751a0. PMID 8264799.
- ↑ Shapiro R (2001). "Cytoplasmic ribonuclease inhibitor". Meth. Enzymol. 341: 611–28. doi:10.1016/S0076-6879(01)41180-3. PMID 11582809.
Further reading
- Kobe B, Deisenhofer J (March 1995). "A structural basis of the interactions between leucine-rich repeats and protein ligands". Nature 374 (6518): 183–6. doi:10.1038/374183a0. PMID 7877692.
- Kobe B, Deisenhofer J (December 1996). "Mechanism of ribonuclease inhibition by ribonuclease inhibitor protein based on the crystal structure of its complex with ribonuclease A". J. Mol. Biol. 264 (5): 1028–43. doi:10.1006/jmbi.1996.0694. PMID 9000628.
- Papageorgiou AC, Shapiro R, Acharya KR (September 1997). "Molecular recognition of human angiogenin by placental ribonuclease inhibitor--an X-ray crystallographic study at 2.0 A resolution". EMBO J. 16 (17): 5162–77. doi:10.1093/emboj/16.17.5162. PMC 1170149. PMID 9311977.
- Suzuki M, Saxena SK, Boix E, Prill RJ, Vasandani VM, Ladner JE, Sung C, Youle RJ (March 1999). "Engineering receptor-mediated cytotoxicity into human ribonucleases by steric blockade of inhibitor interaction". Nat. Biotechnol. 17 (3): 265–70. doi:10.1038/7010. PMID 10096294.
- Shapiro R, Ruiz-Gutierrez M, Chen CZ (September 2000). "Analysis of the interactions of human ribonuclease inhibitor with angiogenin and ribonuclease A by mutagenesis: importance of inhibitor residues inside versus outside the C-terminal "hot spot"". J. Mol. Biol. 302 (2): 497–519. doi:10.1006/jmbi.2000.4075. PMID 10970748.
- Bretscher LE, Abel RL, Raines RT (April 2000). "A ribonuclease A variant with low catalytic activity but high cytotoxicity". J. Biol. Chem. 275 (14): 9893–6. doi:10.1074/jbc.275.14.9893. PMID 10744660.
- Yakovlev G. I., Mitkevich V. A. & Makarov A. A. (2006). Ribonuclease inhibitors 40 (6). pp. 867–874. doi:10.1134/S0026893306060045. Unknown parameter
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