All cellular processes are the results of synchronized actions of several intracellular biochemical pathways. Recent emphasis is to visualize such pathways using appropriate small molecular reagents, dye-labeled proteins, and genetically encoded fluorescent biosensors that produce a luminescence ON response either on selective binding or on reacting with an analyte that is produced through a specific biochemical/ enzymatic transformation. Studying such enzymatic processes by probing the fluorescence response as the read-out signal is expected to provide important insights into crucial biochemical transformations induced by an enzyme in its native form. Many of such studies are extended for monitoring enzymatic transformations under in vitro or in vivo condition. A few of the recent reports reveal that such molecular probes are even capable of quantifying abnormal levels of enzymes in real-time and is linked to the key area of clinical diagnostics and chemical biology. A synchronized analysis of all such reports helps in developing a rationale for designing purpose-built molecular probes or chemodosimeters as well as newer reagents for studying crucial enzymatic process or quantification of the respective enzyme. In this review, an attempt will be there to highlight several recent bioimaging reagents and studies that have provided insights into crucial biochemical or enzymatic transformations.
It is known that reactive oxygen (ROS) and nitrogen (RNS) species play a diverse role in various biological processes, such as inflammation, signal transduction, and neurodegenerative injury, apart from causing various diseases caused by oxidative and nitrosative stresses, respectively, by ROS and RNS. Thus, it is very important to quantify the concentration level of ROS and RNS in live cells, tissues, and organisms. Various smallmolecule-based fluorescent/chemodosimetric probes are reported to quantify and map the effective distribution of ROS/RNS under in vitro/in vivo conditions with a great spatial and temporal resolution. Such reagents are now appreciated as an excellent tool for aiding breakthroughs in modern redox biology. This minireview is a brief, but all-inclusive, account of such molecular probes that have been developed recently.
Nucleoside
polyphosphates (NPPs) are mainly produced in mitochondria
and used as a universal energy source for various cellular events.
Although numerous fluorescent probes for adenosine triphosphate (ATP)
have been reported, they are not ideally suited for live monitoring
of the subtle variation of the mitochondrial ATP level. A new coumarin-based
fluorescent probe is synthesized, and this reagent is utilized for
specific recognition of NPPs in mitochondria by super-resolution microscopy
in physiological condition. Detailed 31P NMR studies reveal
that the probe, L·Zn(II), binds to NPPs through
their pendent phosphate functionalities. Such binding leads to a substantial
enhancement in the luminescence intensity of L·Zn(II)
+ ADP or L·Zn(II) + ATP as compared to L·Zn(II). Thisas well as 1H NMR spectroscopyhas
enabled us to evaluate the probe’s binding affinities to these
NPPs. Structured illumination and wide field fluorescence microscopy
confirmed that this physiologically benign reagent is localized within
mitochondria of live RAW 264.7 macrophage cells. This reagent has
been utilized to probe real-time changes in ATP concentrations within
mitochondria during drug-induced apoptosis.
Super-resolution microscopy (SRM) has revolutionized cell biology, enabling visualization of cellular structures with nanometric resolution, single-molecule sensitivity, and with multiple colors. Here we review how nanocontainers have been used to enhance these techniques.
Contorted polycyclic aromatic hydrocarbons (PAHs), CPA1–2 and CPB1–2, bearing peripheral
five-membered
rings were synthesized employing a palladium-catalyzed cyclopentannulation
reaction using specially designed diaryl acetylene synthons TPE and TPEN with commercially available dibromo-
anthracene DBA and bianthracene DBBA derivatives.
The resulting target compounds CPA1–2 and CPB1–2 were isolated in excellent yield and found to
be highly soluble in common organic solvents, which allowed for their
structural characterization and investigation of the photophysical
properties, disclosing their aggregation-induced emission (AIE) properties
in THF at selective concentration ranges of water fractions in the
solvent mixture. Examination of the contorted PAH structures by means
of density functional theory (DFT) revealed higher electronic conjugation
in the more rigid and planar anthracene-containing CPA1–2 derivatives when compared to the twisted bianthracene-bearing moieties CBPA1–2 with HOMO–LUMO bandgaps (ΔE) of ∼2.32 eV for the former PAHs and ∼2.78
eV for the latter ones.
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