Multiple spectroscopic and computational studies on binding interaction of 2-phenylamino-4-phenoxyquinoline derivatives with bovine serum albumin.

Binding characteristics of potent non-nucleoside HIV-1 reverse transcriptase inhibitors, 4-(2',6'-dimethyl-4'-formylphenoxy)-2-(5″-cyanopyridin-2″ylamino) quinoline (1) and 4-(2',6'-dimethyl-4'-cyanophenoxy)-2-(5″-cyanopyridin-2″ylamino) quinoline (2), to bovine serum albumin (BSA) under simulative physiological conditions were investigated by multiple spectroscopic and computational methods. The experimental results demonstrated that (1) and (2) bound to BSA at site III (subdomain IB), and quenched BSA fluorescence through a static quenching process. The binding interaction of (1) or (2) to BSA forms stable complexes with the binding constants (K) at the level of 10 L/mol and the number of binding site was determined to be 1 for both systems, indicating that new synthesized compounds occupied one site in BSA with moderate binding affinities.

Selective protein photocleavage by fluorescein derivatives.

Modification of the structure of small molecular probe which can act as photocleavage reagent has become a considerable challenge to improve the ability to target specific sites on a large protein. These photoreagents can provide valuable information on the binding site recognition and the mechanism of the photocleavage reaction under photochemical control. In this study, site specific photocleavage of lysozyme and avidin by fluorescein derivatives, fluorescein sodium salt (F-1) and 5(6)-carboxyfluorescein diacetate (F-2) were reported here for the first time.

Tuning the chain length of new pyrene derivatives for site-selective photocleavage of avidin.

Rational design of photoreagents with systematic modifications of their structures can provide valuable information for a better understanding of the protein photocleavage mechanism by these reagents. Variation of the length of the linker connecting the photoactive moiety with the protein anchoring-group allowed us to investigate the control of the protein photocleavage site. A series of new photochemical reagents (PMA-1A, PMA-2A and PMA-3A) with increasing chain lengths is examined in the current study.

Photochemistry and mechanism of designed pyrenyl probe towards promoted cleavage of proteins.

A new photochemical reagent, succinic acid-1(1-pyrene)methylamide (PMA-SUC), was developed to recognize the specific binding sites on model proteins, egg-white lysozyme and avidin. The interaction of the photochemical reagent with the proteins was studied by UV-Vis, fluorescence spectroscopic methods and docking description. PMA-SUC was found to bind to lysozyme and avidin with binding constants (K) of 2.

Use of a molybdenum(VI) complex as artificial protease in protein photocleavage.

In this study, a molybdenum(VI) peroxo α-amino acid complex, MoO(O2)2(α-leucine) (H2O), was prepared and used as an artificial protease for site-specific cleavage of porcine pepsin, a model protein. Cleavage of pepsin by MoO(O2)2(α-leucine) (H2O) was achieved under photochemical conditions at room temperature and pH 7.0.

Selective cleavage of pepsin by molybdenum metallopeptidase.

In this study, the cleavage of protein by molybdenum cluster is reported for the first time. The protein target used is porcine pepsin. The data presented in this study show that pepsin is cleaved to at least three fragments with molecular weights of ∼23, ∼19 and ∼16 kDa when the mixture of the protein and ammonium heptamolybdate tetrahydrate ((NH(4))(6)Mo(7)O(24)·4H(2)O) was incubated at 37°C for 24h.

Photocleavage of avidin by a new pyrenyl probe.

In this study, a new small-molecule-based reagent was designed to recognize and bind to specific site in protein. A new pyrenyl probe, d-biotinyl-1(1-pyrene)methylamide (Py-biotin) was designed and synthesized by coupling of d-biotin to 1(1-pyrene)methylamine hydrochloride. Binding studies and site-specific photocleavage of avidin by Py-biotin were demonstrated.

Chiral protein scissors activated by light: recognition and protein photocleavage by a new pyrenyl probe.

Strong chiral discrimination and site-selective photocleavage of two model proteins, lysozyme and bovine serum albumin (BSA), by new pyrenyl probes are reported here. The enantiomeric pyrenyl probes D-phenylalanine-1(1-pyrene)methylamide (PMA- D-Phe) and L-phenylalanine-1(1-pyrene)methylamide (PMA- l-Phe) were synthesized by coupling the carboxyl function of D-phenylalanine or L-phenylalanine with the amino group of 1(1-pyrene)methylamine. Binding affinities of the two enantiomers with the proteins were quantitated in absorption titrations.

Chiral protein scissors: high enantiomeric selectivity for binding and its effect on protein photocleavage efficiency and specificity.

Chiral recognition of protein-binding sites by a simple organic molecule with selectivities >100 is reported here. The l-isomer of 4(1-pyrene)-3-butyroyl-phenylalanine amide (Py-L-Phe) binds to BSA with an affinity constant (K(b)) of 3 x 10(7) M(-1), whereas the corresponding D-isomer (Py-D-Phe) binds 100 times weaker. The enantiomers showed contrasting spectral changes when bound to BSA.