Interaction of flavonoids with serum albumin: A review

: Flavonoids are plant products abundant in every day diet and claimed to be beneficial for human health. After absorption, flavonoids are transported by the serum albumin (SA), the most abundant carrier blood protein, through formation of flavonoids-SA complex. This review deals with the current state of knowledge on flavonoids-SA complex over the past 10 years, mainly involved multi-spectroscopic techniques and molecular dynamics simulation studies to explore the binding mechanism, thermodynamics and structural aspects of flavonoids binding to SA. Especially, the novel method, capillary electrophoresis, high performance affinity chromatography approach, native mass spectrometry and microscale thermophoresis used in characterization of the interaction between flavonoids and SA as well as flavonoid-based fluorescent probe for SA measurement are also included in this review.

Characterization, Expression Profiling, and Functional Analyses of a 4CL-Like Gene of Populus trichocarpa

Adenosine 5′-monophosphate (AMP) (adenylate)-forming acetyl-CoA synthetase (ACS) catalyzes the formation of acetyl-coenzyme A (CoA), and the ACS family is closely related to the 4-coumarate CoA ligase (4CL) family. In this study, a 4CL-like gene was cloned from Populus trichocarpa and named Pt4CL-like. Characterization of Pt4CL-like, using bioinformatics, showed that it contained box I and box II domains at the end of the C-terminal sequence, and there is a characteristic sequence of ACS, namely, peroxisome-targeting sequence (PTS). Real-time PCR results showed that the 4CL-like gene was expressed in all tissues tested, and was highly expressed in the stems. A denaturation and renaturation process was conducted, and the recombinant Pt4CL-like protein was purified through HisTrapTM high performance affinity chromatography. It showed Pt4CL-like protein did not catalyze substrates of 4CL, but could significantly catalyzed sodium acetate. These results indicate that Pt4CL-like protein belongs to the ACS family, providing a theoretical basis for further analysis and comparison of the functions of adenylate-forming enzymes and 4CL family.

Aspects of Drug-Protein Binding and Methods of Analyzing the Phenomenon

In recent decades, drug-protein interactions have been widely studied and several methods of analysis of these phenomena have been developed and improved. These can be classified into separation, physical, chromatographic and electrophoretic methods. This review depicts the assumptions and mechanisms of methods from each group, details their strengths and weaknesses, and presents examples of their usage from the literature. Equilibrium dialysis, ultrafiltration, Hummel-Dreyer method or high performance affinity chromatography are given as representative examples, but this issue is far more expanded. Nowadays, increasing attention is paid to the computational methods and molecular modeling which are convenient tools to estimate protein binding affinity based on the physicochemical properties of compounds. To gain a broader overview, the study also examines the protein binding ability and pharmacotherapy of drugs against a range of substrates such as plasma, skin, tissue and human milk.

High performance affinity chromatography and related separation methods for the analysis of biological and pharmaceutical agents

The last few decades have witnessed the development of many high-performance separation methods that use biologically related binding agents.