Advertisehere

BD Biosciences High Quality Reagents banner

Latest promotion FROM:

BD (Becton, Dickinson and Company)

View promotion

$Rce1 (Prenyl protein protease RCE1B,\nfarnesylated protein-converting enzyme 2)$

Supplier:

United States Biological

Cat no: R1250-05

Rce1 (Prenyl protein protease RCE1B,\nfarnesylated protein-converting enzyme 2)

Protein prenylation plays a vital role in the membrane localization and function of some proteins. Proteins containing C-terminal CAAX sequence motifs undergo several posttranslational processing steps. First, the cysteine is modified by specific transferases that covalently attach prenyl lipids, either a 15-carbon farnesyl or a 20-carbon geranylgeranyl isoprenyl lipid. A specific protease then cleaves the AAX tripeptide from the protein, leaving the prenylated cysteine as the new C terminus. The final modification is methylation of the carboxyl group of the now C-terminal prenylcysteine.\n\nA genetic screen in S. cerevisiae resulted in the identification of a candidate gene for a prenyl protein protease, RCE1 (Boyartchuk et al., 1997). In a database search, Otto et al. (1999) identified a human EST as a potential homolog of yeast RCE1. A full-length RCE1 cDNA was assembled from clones isolated from a human fetal brain cDNA library and a HUVEC cDNA library. RCE1 encodes a deduced 329-amino acid protein that shares 42% identity over 105 amino acids with the yeast protein and contains several predicted transmembrane domains. By Northern blot analysis, Otto et al. (1999) detected a 1.8-kb RCE1 transcript in heart, brain, placenta, lung, liver, muscle, kidney, and pancreas. A weaker 2.4-kb transcript was also detected in some tissues. Northern blot analysis of mouse tissues revealed broad expression and multiple transcript sizes. Otto et al. (1999) concluded that these transcripts may represent alternatively spliced variants of RCE1.\n\nFreije et al. (1999) identified RCE1, which they called FACE2, by EST database searching for sequences with similarity to yeast Rce1, a protein essential for the proteolytic processing of yeast farnesylated/prenylated proteins. They cloned a human RCE1 cDNA from an ovary cDNA library and found that it encodes a deduced 329-amino acid protein with an HXXE motif characteristic of some metalloproteinases. Freije et al. (1999) identified regions of high hydrophobicity, consistent with RCE1 being a polytopic integral membrane protein. Using in vitro translation product for SDS-PAGE analysis, they detected a 30kD RCE1 protein. Northern blot analysis detected ubiquitous expression of a 1.4-kb RCE1 transcript.\n\nOtto et al. (1999) detected a high level of prenyl protease activity with both yeast and human RCE1 proteins expressed in a recombinant baculovirus system. The partial recombinant human RCE1 clone used all forms of Ras as substrates, suggesting that the enzyme can use a broad range of CAAX-type prenyl protein substrates. By competition studies, Otto et al. (1999) demonstrated that the protease activity of human RCE1 activity is specific for prenylated proteins and that RCE1 recognizes short prenylated peptides.\n\nBy radiation hybrid analysis and FISH, Freije et al. (1999) mapped the RCE1 gene to chromosome 11q13, a region frequently amplified in carcinomas and lymphomas.\n\nApplications: \nSuitable for use in Western Blot,. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1::500\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. \n\nQuality Control Testing\nImmunoblot Analysis: A 1:500 dilution of this lot detected endogenous Rce1 in lysate from mouse heart. Immunoblot Analysis Lysate from mouse heart was resolved by electrophoresis, transferred to PVDF, and probed with anti-Rce1 for 2 hours at room temperature. Proteins were visualized via HRP and chemiluminescent detection. Arrow indicates Rce1.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

R1250-05

Size

200ul

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Mouse

Form

Supplied as a liquid in 0.02M PBS, 0.25M sodium chloride containing 0.1% sodium azide and 50% glycerol.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Plummer, Lisa J, et al (2006). Mutational analysis of the ras converting enzyme reveals a requirement for glutamate and histidine residues. J Biol Chem 281: 4596-605.\n2. Hollander, Irwin J, et al (2003). Human Ras converting enzyme endoproteolytic specificity at the P2' and P3' positions of K-Ras-derived peptides. Biochim Biophys Acta 1649: 24-9.\n3. Maske, Christopher P, et al (2003). A carboxyl-terminal interaction of lamin B1 is dependent on the CAAX endoprotease Rce1 and carboxymethylation. J Cell Biol 162: 1223-32.\nGeneral References:\n1. Boyartchuk, V. L.; Ashby, M. N.; Rine, J.: Modulation of Ras and a-factor function by carboxyl-terminal proteolysis. Science 275: 1796-1800, 1997. PubMed ID : 9065405\n2. Freije, J. M. P.; Blay, P.; Pendas, A. M.; Cadinanos, J.; Crespo, P.; Lopez-Otin, C.: Identification and chromosomal location of two human genes encoding enzymes potentially involved in proteolytic maturation of farnesylated proteins. Genomics 58: 270-280, 1999. PubMed ID : 10373325\n3. Otto, J. C.; Kim, E.; Young, S. G.; Casey, P. J.: Cloning and characterization of a mammalian prenyl protein-specific protease. J. Biol. Chem. 274: 8379-8382, 1999. PubMed ID : 10085068\n