PMID: 3890941Mar 26, 1985Paper

Limited proteolysis of thermolysin by subtilisin: isolation and characterization of a partially active enzyme derivative

Biochemistry
C VitaA Fontana

Abstract

Incubation of the neutral metalloendopeptidase thermolysin at pH 9-10 in the presence of 10 mM CaCl2 for 2 days at room temperature with subtilisin at a 50:1 molar ratio leads to a derivative possessing lower (approximately 3%) but intrinsic catalytic activity. This derivative, called thermolysin S, was isolated by gel filtration in approximately 80% yield and then separated from some residual intact thermolysin by an affinity chromatographic step on Sepharose-Gly-D-Phe. It was found that thermolysin S results from a tight association of two polypeptide fragments of apparent Mr of 24000 and 10000. Dissociation of the complex was achieved under strong denaturing conditions, such as gel filtration on a column equilibrated and eluted with 5 M guanidine hydrochloride. The positions of the clip sites were defined by amino acid analysis, end-group determination, and amino acid sequencing of the isolated fragments and shown to lie between Thr-4 and Ser-5, between Thr-224 and Gln-225, and also between Gln-225 and Asp-226. Thermolysin S, which is therefore a stable complex of fragments 5-224(225) and 225(226)-316, shows a shift in optimum pH of about 1 unit toward the acid range with respect to intact thermolysin and a Km essentially un...Continue Reading

References

Jan 1, 1975·Advances in Immunology·M Reichlin
Jan 1, 1975·Advances in Protein Chemistry·C B Anfinsen, H A Scheraga
Sep 1, 1975·Analytical Biochemistry·E Mendez, C Y Lai
Nov 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·K TitaniE H Fischer
Jan 1, 1978·CRC Critical Reviews in Biochemistry·R S Roche, G Voordouw
Sep 1, 1977·The Biochemical Journal·A FontanaF M Veronese
Mar 9, 1976·Biochemistry·F W DahlquistW L Bigbee
Dec 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·D H SachsC B Anfinsen
Sep 5, 1973·Biochemical and Biophysical Research Communications·M K PangburnH Neurath
Jan 1, 1974·Methods in Enzymology·K A WalshM K Pangburn
Jan 1, 1974·CRC Critical Reviews in Biochemistry·E H Strickland
Jan 1, 1974·Biochemistry·Y BursteinH Neurath
Oct 14, 1972·Journal of Molecular Biology·P M ColmanB W Matthews
Oct 8, 1969·Biochemical and Biophysical Research Communications·S A LattB L Vallee
Jan 1, 1967·Biochemistry·M S Doscher, C H Hirs
Mar 4, 1967·Nature·G KarthaD Harker
Jan 1, 1983·International Journal of Peptide and Protein Research·C VitaA Fontana
Jan 1, 1981·CRC Critical Reviews in Biochemistry·I M Chaiken
Oct 5, 1982·Journal of Molecular Biology·M A Holmes, B W Matthews
Mar 3, 1981·Biochemistry·A Dautry-Varsat, J R Garel
Jan 1, 1948·Acta Pathologica Et Microbiologica Scandinavica·O OUCHTERLONY
Oct 1, 1980·Biophysical Journal·P S KimR L Baldwin

❮ Previous
Next ❯

Citations

Apr 1, 1997·Protein Science : a Publication of the Protein Society·P Polverino de LauretoA Fontana
Jan 1, 1987·The International Journal of Biochemistry·D A CowanH W Morgan
Jan 1, 1987·Progress in Biophysics and Molecular Biology·R Jaenicke
Apr 16, 1998·Biochimica Et Biophysica Acta·S J Hubbard
Jun 19, 2002·Protein Science : a Publication of the Protein Society·Chung-Jung TsaiRuth Nussinov
Apr 15, 1986·European Journal of Biochemistry·G FassinaA Fontana
Aug 15, 1989·European Journal of Biochemistry·C VitaR Jaenicke
Dec 31, 1985·Biochemical and Biophysical Research Communications·L T TamE G Erdös
Sep 17, 2014·Journal of Basic Microbiology·Irfana IqbalIkram Ul-Haq
Jan 1, 1997·Folding & Design·A FontanaM Zambonin
Feb 28, 1990·Biochemical and Biophysical Research Communications·M BianchiM A Coletti-Previero
Oct 7, 2005·BioTechniques·Rimantas Sapranauskas, Arvydas Lubys
Dec 1, 1986·International Journal of Peptide and Protein Research·R J Corbett, R S Roche
Aug 1, 1986·The American Journal of Physiology·R J Beynon, J S Bond
Jul 1, 1988·Archives of Biochemistry and Biophysics·S B KurlandskyH R Levy

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.