Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

To build an enzyme....

J R Knowles1

  • 1Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.

Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
|May 29, 1991
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Antimicrobial action of carvacrol at different stages of dual-species biofilm development by Staphylococcus aureus and Salmonella enterica serovar Typhimurium.

Applied and environmental microbiology·2005
Same author

New Construction Act.

Health estate·1998
Same author

East meets West: a comparison of eastern block/western aeromedical practices.

Aviation, space, and environmental medicine·1997
Same author

Introns and the origin of protein-coding genes.

Science (New York, N.Y.)·1995
Same author

Complementation of fragments of triosephosphate isomerase defined by exon boundaries.

Biochemistry·1995
Same author

Phosphoglycerate kinase and triosephosphate isomerase from the hyperthermophilic bacterium Thermotoga maritima form a covalent bifunctional enzyme complex.

The EMBO journal·1995
Same journal

The microlandscapes of tree trunks: the effect of lichen and tree-level characteristics on arthropod communities.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
Same journal

Centimetre-scale landscapes to assess the motion behaviour and cognition of gastropods and bivalves.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
Same journal

Intertidal microcosms of wave-swept rocky shores: ecological and physiological insights from a uniquely stressful environment.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
Same journal

Temporal and spatial variation in temperature and oxygen at the microscale: key niche axes for aquatic life.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
Same journal

Natural microcosms in ecology: fulfilling the promise of model systems?

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
Same journal

Microbe-induced galls and plant defence: metabolite crosstalk in a co-evolutionary battle.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences·2026
See all related articles

Triosephosphate isomerase uses specific structural features, including alpha-helices and a flexible loop, to bind substrates and catalyze reactions efficiently. These elements facilitate substrate binding, enolization, and prevent intermediate loss for optimal enzyme function.

Area of Science:

  • Biochemistry
  • Enzymology
  • Structural Biology

Background:

  • Enzyme catalysis relies on precise structural components for function.
  • Triosephosphate isomerase (TIM) is a model enzyme for studying catalytic mechanisms.

Purpose of the Study:

  • To elucidate the structural determinants of triosephosphate isomerase function.
  • To understand the roles of specific amino acid residues and protein structures in catalysis.

Main Methods:

  • Analysis of hydrogen bonding for substrate recognition and binding.
  • Identification of catalytic residues (base and general acid) using mechanistic principles.
  • Spectroscopic techniques (Fourier transform infrared and 15N NMR) to determine imidazole protonation state.
  • Examination of protein dynamics, specifically flexible loops, for intermediate sequestration.

Related Experiment Videos

Main Results:

  • Substrate phospho group binding involves four main-chain hydrogen bonds, with two from an alpha-helix.
  • Catalysis requires a carboxylate base and a neutral imidazole general acid.
  • The imidazole's pKa is lowered to <4.5 due to its position on an alpha-helix.
  • A flexible loop sequesters the reaction intermediate, enhancing catalytic efficiency.

Conclusions:

  • Specific structural arrangements, including alpha-helices and a flexible loop, are critical for triosephosphate isomerase activity.
  • The enzyme utilizes a unique protonation state for its general acid catalyst.
  • Structural features optimize substrate binding, catalysis, and intermediate stabilization, leading to high efficiency.