Structure of bovine pancreatic cholesterol esterase at 1.6 A: novel structural features involved in lipase activation

Biochemistry
J C ChenR M Stroud

Abstract

The structure of pancreatic cholesterol esterase, an enzyme that hydrolyzes a wide variety of dietary lipids, mediates the absorption of cholesterol esters, and is dependent on bile salts for optimal activity, is determined to 1.6 A resolution. A full-length construct, mutated to eliminate two N-linked glycosylation sites (N187Q/N361Q), was expressed in HEK 293 cells. Enzymatic activity assays show that the purified, recombinant, mutant enzyme has activity identical to that of the native, glycosylated enzyme purified from bovine pancreas. The mutant enzyme is monomeric and exhibits improved homogeneity which aided in the growth of well-diffracting crystals. Crystals of the mutant enzyme grew in space group C2, with the following cell dimensions: a = 100.42 A, b = 54.25 A, c = 106.34 A, and beta = 104.12 degrees, with a monomer in the asymmetric unit. The high-resolution crystal structure of bovine pancreatic cholesterol esterase (Rcryst = 21.1%; Rfree = 25.0% to 1.6 A resolution) shows an alpha-beta hydrolase fold with an unusual active site environment around the catalytic triad. The hydrophobic C terminus of the protein is lodged in the active site, diverting the oxyanion hole away from the productive binding site and the cat...Continue Reading

References

Sep 25, 1992·Nucleic Acids Research·L Good, R N Nazar
Jan 1, 1994·Protein Science : a Publication of the Protein Society·P GrochulskiM Cygler
Sep 15, 1993·European Journal of Biochemistry·T SugoD Lombardo
Jun 20, 1993·Journal of Molecular Biology·R A LaskowskiJ M Thornton

❮ Previous
Next ❯

Citations

Dec 22, 1999·Lipids·T TsujitaH Okuda
May 7, 2013·Chemico-biological Interactions·Yingling WeiJiale Huang
May 17, 2005·Methods : a Companion to Methods in Enzymology·Dean Gilham, Richard Lehner
Dec 12, 2001·Biochimica Et Biophysica Acta·R E SmithD M Yourtee
Dec 23, 1999·Current Opinion in Structural Biology·M Nardini, B W Dijkstra
Jul 20, 2002·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Dietmar E BreithauptUrsula Wirt
Jul 13, 2005·Natural Product Reports·Gerard P McGlacken, Ian J S Fairlamb
Feb 16, 2002·European Journal of Biochemistry·Susanne LindquistOlle Hernell
Oct 29, 1999·European Journal of Biochemistry·K M LoomesA M Roberton
Feb 6, 2009·Molecular Biology and Evolution·Simon G Williams, Simon C Lovell
Oct 25, 2000·Protein Science : a Publication of the Protein Society·S TerzyanX C Zhang
Apr 5, 2001·Journal of Molecular Biology·A E ToddJ M Thornton
Feb 20, 2003·Proceedings of the National Academy of Sciences of the United States of America·Natalia FriedlandAnn M Stock
Nov 25, 2011·Enzyme and Microbial Technology·Naoya MorinagaAkio Sugihara
Jun 24, 2008·Biochimica Et Biophysica Acta·Sylvie FernandezVincent Jannin
May 24, 2008·Journal of Bioscience and Bioengineering·Atsushi MaedaAkio Sugihara
Dec 8, 2007·Biosensors & Bioelectronics·Sunil K AryaBansi D Malhotra
Nov 13, 2012·Journal of Clinical Laboratory Analysis·Pornpen SrisawasdiMartin H Kroll
Dec 13, 2006·Biochimica Et Biophysica Acta·Hongyu XiangNaoki Takaya
Nov 19, 2011·Bioengineered Bugs·Roy D Sleator
May 4, 2013·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Dafeng ChuThomas Kissel
Nov 15, 2011·Bioorganic & Medicinal Chemistry·Sabrina HengAndrew D Abell
Jul 15, 1999·Structure·P HeikinheimoD L Ollis
Jan 21, 2017·Chembiochem : a European Journal of Chemical Biology·Thomas DriantEtienne Derat

❮ 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.