An emerging paradigm in enzymology is that transient high-energy structural states play crucial roles in enzymatic reaction cycles. Generally, these high-energy or 'invisible' states cannot be studied directly at atomic resolution using existing structural and spectroscopic techniques owing to their low populations or short residence times. Here we report the direct NMR-based detection of the m...

Elucidating the origin of enzymatic catalysis stands as one the great challenges of contemporary biochemistry and biophysics. The recent emergence of computational enzymology has enhanced our atomistic-level description of biocatalysis as well the kinetic and thermodynamic properties of their mechanisms. There exists a diversity of computational methods allowing the investigation of specific en...

Levi-Montalcini, R. & Angeletti, P. U. (1962). Annu. Rev. Physiol. 24, 11. Mannik, M. & Kunkel, H. G. (1963). J. exp. Med. 117, 213. Markert, C. L. & Hunter, R. L. (1959). J. Histochem. Cytochem. 7, 42. Markert, C. L. & M6ller, F. (1959). Proc. nat. Acad. Sci., Wash., 45, 753. Ornstein, L. (1962). Disc Electrophoresis. Rochester,N.Y.: Eastman Kodak Co. Ouchterlony, 0. (1953). Acta path. microbi...

My understanding is that “reflections” can cover a multitude of sins. I have previously written a brief history of my career, and do not want to duplicate that exposition (1). Instead, this “reflection” will trace a primary focus of my laboratory for many decades, namely how do enzymes achieve their remarkable catalytic efficiency? (Indeed this has been a focal point of the field of biochemistr...

The glpX gene from Francisella tularensis encodes for the class II fructose 1,6-bisphosphatase (FBPaseII) enzyme. The glpX gene has been verified to be essential in F. tularensis, and the inactivation of this gene leads to impaired bacterial growth on gluconeogenic substrates. In the present work, we have complemented a ∆glpX mutant of Escherichia coli with the glpX gene of F. tularensis (FTF16...

Robin van der Lee,*,†,‡ Marija Buljan,†,▲ Benjamin Lang,†,▲ Robert J. Weatheritt,†,▲ Gary W. Daughdrill, A. Keith Dunker, Monika Fuxreiter, Julian Gough, Joerg Gsponer, David T. Jones, Philip M. Kim, Richard W. Kriwacki, Christopher J. Oldfield, Rohit V. Pappu, Peter Tompa, Vladimir N. Uversky, Peter E. Wright, and M. Madan Babu*,† †MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cam...

Enzyme inhibition due to the reversible binding of reaction products is common and underlies the origins of negative feedback inhibition in many metabolic and signaling pathways. Product inhibition generates non-linearity in steady-state time courses of enzyme activity, which limits the utility of well-established enzymology approaches developed under the assumption of irreversible product rele...

Description: The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today truly an essential publication for researchers in all fields ...

Laura Fancellu, MD Giovanni Andrea Deiana, PhD GianPietro Sechi, MD A 41-year-old man with chronic hypertension presented in a transient, global confusional state. The patient had neuropathic pain and kidney disease, the significance not recognized by his doctors. Family history was positive for hypertension and kidney disease. Brain CT, MRI, and magnetic resonance angiography showed abnormalit...

Diacylglycerol kinases (DGKs) are a family of enzymes that catalyze the ATP-dependent phosphorylation of diacylglycerol (DAG) to phosphatidic acid (PtdOH). The recognition of the importance of these enzymes has been increasing ever since it was determined that they played a role in the phosphatidylinositol (PtdIns) cycle and a number of excellent reviews have already been written [(see van Blit...