Demonstration of two distinct high molecular weight proteases in rabbit reticulocytes, one of which degrades ubiquitin conjugates - PubMed
- ️Thu Jan 01 1987
. 1987 Feb 25;262(6):2451-7.
- PMID: 3029081
Free article
Demonstration of two distinct high molecular weight proteases in rabbit reticulocytes, one of which degrades ubiquitin conjugates
L Waxman et al. J Biol Chem. 1987.
Free article
Abstract
Reticulocytes contain a nonlysosomal proteolytic pathway that requires ATP and ubiquitin. By DEAE chromatography and gel filtration, we were able to fractionate the ATP-dependent system into a 30-300-kDa fraction that catalyzes the ATP-dependent conjugation of ubiquitin to substrates ("Conjugation Fraction") and a high mass fraction (greater than 450 kDa) necessary for hydrolysis of the conjugated proteins. The latter contains two distinct proteases. One protease is unusually large, approximately 1500 kDa, and degrades proteins only when ATP and the conjugating fractions are added. This activity precipitates at 0-38% (NH4)2SO4 saturation and is essential for ATP-dependent proteolysis. Like crude extracts, it is labile in the absence of nucleotides and is inhibited by heparin, poly(Glu-Ala-Tyr), 3,4-dichloroisocoumarin, hemin, decavanadate, N-ethylmaleimide, and various peptide chloromethyl ketones. It lacks amino-peptidase and insulin-degrading activities and does not require tRNA for activity. The ubiquitin-conjugate degrading enzyme, which we suggest be named UCDEN, is inactive against substrates that cannot undergo ubiquitin conjugation. The smaller protease (670 kDa), which precipitates at 40-80% (NH4)2SO4 saturation, does not require ATP or ubiquitin and is therefore not required for ATP-dependent proteolysis. It is stimulated by N-ethylmaleimide and 3,4-dichloroisocoumarin and is stable at 37 degrees C. It hydrolyzes fluorometric tetrapeptides and proteins, including proteins which cannot be conjugated to ubiquitin. Thus, reticulocytes contain two large cytosolic proteases: one is essential for the degradation of ubiquitin conjugates, while the function of the other is uncertain.
Similar articles
-
ATP-dependent proteolysis and the role of ubiquitin in rabbit cardiac muscle.
Gehrke PP, Jennissen HP. Gehrke PP, et al. Biol Chem Hoppe Seyler. 1987 Jun;368(6):691-708. doi: 10.1515/bchm3.1987.368.1.691. Biol Chem Hoppe Seyler. 1987. PMID: 3040036
-
Protease/inhibitor mechanisms involved in ATP-dependent proteolysis.
Etlinger JD, Gu M, Li X, Weitman D, Rieder RF. Etlinger JD, et al. Revis Biol Celular. 1989;20:197-216. Revis Biol Celular. 1989. PMID: 2561542 Review.
-
Degradation of proteins by the ubiquitin-mediated proteolytic pathway.
Ciechanover A, Gonen H, Elias S, Mayer A. Ciechanover A, et al. New Biol. 1990 Mar;2(3):227-34. New Biol. 1990. PMID: 2177651 Review.
Cited by
-
The roles of the proteasome pathway in signal transduction and neurodegenerative diseases.
Chen JJ, Lin F, Qin ZH. Chen JJ, et al. Neurosci Bull. 2008 Jun;24(3):183-94. doi: 10.1007/s12264-008-0183-6. Neurosci Bull. 2008. PMID: 18500392 Free PMC article. Review.
-
Ciechanover A. Ciechanover A. Rambam Maimonides Med J. 2012 Jan 31;3(1):e0001. doi: 10.5041/RMMJ.10068. Print 2012 Jan. Rambam Maimonides Med J. 2012. PMID: 23908826 Free PMC article.
-
Benoist P, Müller A, Diem HG, Schwencke J. Benoist P, et al. J Bacteriol. 1992 Mar;174(5):1495-504. doi: 10.1128/jb.174.5.1495-1504.1992. J Bacteriol. 1992. PMID: 1537794 Free PMC article.
-
Reversible phosphorylation of the 26S proteasome.
Guo X, Huang X, Chen MJ. Guo X, et al. Protein Cell. 2017 Apr;8(4):255-272. doi: 10.1007/s13238-017-0382-x. Epub 2017 Mar 3. Protein Cell. 2017. PMID: 28258412 Free PMC article. Review.
-
The unravelling of the ubiquitin system.
Ciechanover A. Ciechanover A. Nat Rev Mol Cell Biol. 2015 May;16(5):322-4. doi: 10.1038/nrm3982. Nat Rev Mol Cell Biol. 2015. PMID: 25907614 Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources