RNPA E.Coli
Escherichia Coli.
ECK3696, Rnase P protein, RnaseP protein, b3704, JW3681, Ribonuclease P protein component, EC 3.1.26.5, Protein C5.
Greater than 90.0% as determined by SDS-PAGE.
Description
RNPA E.Coli Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 119 amino acids (1-119a.a) and having a molecular mass of 13.7kDa.
RNPA is purified by proprietary chromatographic techniques.
Product Specs
Ribonuclease P (RNase P), specifically its protein component rnpA, is a crucial enzyme found in bacteria, archaea, and eukarya. It partners with the M1 RNA (rnpB) to form a functional ribonucleoprotein complex. This complex is responsible for the maturation of tRNA molecules by cleaving off the 5' leader sequences from precursor tRNAs. Beyond this, RNase P plays a vital role in the processing of specific polycistronic tRNA transcripts like valV valW, leuQ leuP leuV, and secG leuU by separating them. This function is essential due to the dependency of certain tRNAs on RNase P for their release from these polycistronic transcripts.
This product consists of the recombinant RNPA protein derived from Escherichia coli (E. coli). Produced in E. coli, this non-glycosylated protein is a single polypeptide chain with a molecular weight of 13.7 kDa. It encompasses amino acids 1 to 119 of the RNPA sequence. The purification of RNPA is achieved through proprietary chromatographic techniques.
The RNPA protein is provided in a solution with a concentration of 0.25 mg/ml. The solution is buffered with Phosphate-Buffered Saline (pH 7.4) and contains 10% glycerol.
For short-term storage (up to 2-4 weeks), the product can be stored at 4°C. For extended storage, it is recommended to freeze the product at -20°C. The addition of a carrier protein such as HSA or BSA (0.1%) is advisable for long-term storage. It is important to minimize freeze-thaw cycles to maintain product integrity.
The purity of this product is greater than 90.0% as determined by SDS-PAGE analysis.
ECK3696, Rnase P protein, RnaseP protein, b3704, JW3681, Ribonuclease P protein component, EC 3.1.26.5, Protein C5.
Escherichia Coli.
MVKLAFPREL RLLTPSQFTF VFQQPQRAGT PQITILGRLN SLGHPRIGLT VAKKNVRRAH ERNRIKRLTR ESFRLRQHEL PAMDFVVVAK KGVADLDNRA LSEALEKLWR RHCRLARGS
Product Science Overview
Ribonuclease P (RNase P) is a ribonucleoprotein complex that plays a crucial role in the maturation of transfer RNA (tRNA) molecules. It is responsible for the specific cleavage of the 5′ leader sequence from precursor tRNA (pre-tRNA) transcripts, converting them into functional tRNA molecules. This enzyme is ubiquitous, found in all cells of bacteria, archaea, and eukarya .
In Escherichia coli (E. coli), RNase P consists of two main components: a large RNA molecule (P RNA) and a small protein subunit (P protein). The RNA component alone can act as a catalyst in vitro under high concentrations of monovalent and divalent cations, but the protein subunit is essential for physiological activity .
The protein component of RNase P in E. coli, often referred to as the C5 protein, acts as a cofactor for the catalytic M1 RNA subunit. This protein enhances the substrate affinity and specificity of the RNA component, facilitating the efficient processing of pre-tRNA .
To facilitate biochemical and biophysical studies, the protein component of RNase P from various sources, including Bacillus subtilis, has been overproduced in E. coli. This involves using the native amino acid sequence with optimized codons for expression in E. coli. The recombinant protein is then purified using techniques such as cation exchange chromatography .
The recombinant RNase P protein component is characterized by its secondary structure, which includes a combination of α-helix and β-sheet structures. It is quite stable, with a melting temperature (T_m) of 67°C. The identity of the recombinant protein as a cofactor of RNase P is established by its ability to stimulate the activity of the RNA component in low ionic strength buffer in a 1:1 stoichiometry .
The recombinant expression and purification of the RNase P protein component are essential for high-resolution crystallographic analyses and other biochemical studies. These studies help in understanding the structure-function relationship of RNase P and its role in tRNA maturation. Additionally, the methods developed for the purification of the RNase P protein component can be applied to other RNA-binding proteins .
