The interest of biologists in boron (B) has largely been focused on its role in plants for which B was established as essential in 1923 (Warington, 1923[296]). Evidence that B has a biological role in other organisms was first indicated by the establishment of essentiality of B for diatoms (Smyth and Dugger, 1981[296]) and cyanobacteria (Bonilla et al., 1990[296]; Garcia-Gonzalez et al., 1991[296]; Bonilla et al., 1997[296]). Recently, B was shown to stimulate growth in yeast (Bennett et al., 1999[296]) and to be essential for zebrafish (Danio rerio) (Eckhert and Rowe, 1999[296]; Rowe and Eckhert, 1999[296]) and possibly for trout (Oncorhynchus mykiss) (Eckhert, 1998[296]; Rowe et al., 1998[296]), frogs (Xenopus laevis) (Fort et al., 1998[296]) and mouse (Lanoue et al., 2000[296]). There is also preliminary evidence to suggest that B has at least a beneficial role in humans (Nielsen, 2000[296]).

While research into the role of B in plants has been ongoing for 80 years it has only been in the past 5 years that the first function of B in plants has been defined. Boron is now known to be essential for cell wall structure and function, likely through its role as a stabilizer of the cell wall pectic network and subsequent regulation of cell wall pore size. A role for B in plant cell walls, however, is inadequate to explain all of the effects of B deficiency seen in plants. The suggestion that B plays a broader role in biology is supported by the discovery that B is essential for animals where a cellulose-rich cell wall is not present. Careful consideration of the physical and chemical properties of B in biological systems, and of the experimental data from both plants and animals suggests that B plays a critical role in membrane structure and hence function. Verification of B association with membranes would represent an important advance in modern biology.

For several decades there has been uncertainty as to the mechanisms of B uptake and transport within plants. This uncertainty has been driven by a lack of adequate methodology to measure membrane fluxes of B at physiologically relevant concentrations. Recent experimentation provides the first direct measurement of membrane permeability of B and illustrates that passive B permeation contributes sufficient B at adequate levels of B supply, but would be inadequate at conditions of marginal B supply. The hypothesis that an active, carrier mediated process is involved in B uptake at low B supply is supported by research demonstrating that B uptake can be stimulated by B deprivation, that uptake rates follow a Michaelis-Menton kinetics, and can be inhibited by application of metabolic inhibitors. Since the mechanisms of element uptake are generally conserved between species, an understanding of the processes of B uptake is relevant to studies in both plants and animals.

The study of B in plant biology has progressed markedly in the last decade and we are clearly on the cusp of additional, significant discoveries. Research in this field will be greatly stimulated by the discovery that B is essential for animals, a discovery that will not only encourage the participation of a wider cadre of scientists but will refocus the efforts of plant biologists toward a determination of roles for B outside the plant cell wall. Determination of the function of B in biology and of the mechanisms of B uptake in biological systems, is essential to our understanding and management of B deficiency and toxicity in plants and animals in both agricultural and natural environments. Through an analysis of existing data and the development of new hypotheses, this review aims to provide a vision of the future of research into the biology of boron.

Boron in Plant Biology

Proton channels exist in a wide variety of membrane proteins where they transport protons rapidly and efficiently. Usually the proton pathway is formed mainly by water molecules present in the protein, but its function is regulated by titratable groups on critical amino acid residues in the pathway. All proton channels conduct protons by a hydrogen-bonded chain mechanism in which the proton hops from one water or titratable group to the next. Voltage-gated proton channels represent a specific subset of proton channels that have voltage- and time-dependent gating like other ion channels. However, they differ from most ion channels in their extraordinarily high selectivity, tiny conductance, strong temperature and deuterium isotope effects on conductance and gating kinetics, and insensitivity to block by steric occlusion. Gating of H+ channels is regulated tightly by pH and voltage, ensuring that they open only when the electrochemical gradient is outward. Thus they function to extrude acid from cells. H+ch...

https://www.physiology.org/doi/pdf/10.1152/physrev.00028.2002

Voltage-Gated Proton Channels and Other Proton Transfer Pathways

Mixed lineage kinase domain-like protein (MLKL) was identified to function downstream of receptor interacting protein 3 (RIP3) in tumor necrosis factor-α (TNF)-induced necrosis (also called necroptosis). However, how MLKL functions to mediate necroptosis is unknown. By reconstitution of MLKL function in MLKL-knockout cells, we showed that the N-terminus of MLKL is required for its function in necroptosis. The oligomerization of MLKL in TNF-treated cells is essential for necroptosis, as artificially forcing MLKL together by using the hormone-binding domain (HBD*) triggers necroptosis. Notably, forcing together the N-terminal domain (ND) but not the C-terminal kinase domain of MLKL causes necroptosis. Further deletion analysis showed that the four-α-helix bundle of MLKL (1-130 amino acids) is sufficient to trigger necroptosis. Both the HBD*-mediated and TNF-induced complexes of MLKL(ND) or MLKL are tetramers, and translocation of these complexes to lipid rafts of the plasma membrane precedes cell death. The homo-oligomerization is required for MLKL translocation and the signal sequence for plasma membrane location is located in the junction of the first and second α-helices of MLKL. The plasma membrane translocation of MLKL or MLKL(ND) leads to sodium influx, and depletion of sodium from the cell culture medium inhibits necroptosis. All of the above phenomena were not seen in apoptosis. Thus, the MLKL oligomerization leads to translocation of MLKL to lipid rafts of plasma membrane, and the plasma membrane MLKL complex acts either by itself or via other proteins to increase the sodium influx, which increases osmotic pressure, eventually leading to membrane rupture.

/pdf/translocation-of-mixed-lineage-kinase-domain-like-protein-to-321ohzj9wv.pdf

Translocation of mixed lineage kinase domain-like protein to plasma membrane leads to necrotic cell death

There is strong evidence that aquaporins are central components in plant water relations. Plant species possess more aquaporin genes than species from other kingdoms. According to sequence similarities, four major groups have been identified, which can be further divided into subgroups that may correspond to localization and transport selectivity. They may be involved in compatible solute distribution, gas-transfer (CO2, NH3) as well as in micronutrient uptake (boric acid). Recent advances in determining the structure of some aquaporins gives further details on the mechanism of selectivity. Gating behaviour of aquaporins is poorly understood but evidence is mounting that phosphorylation, pH, pCa and osmotic gradients can affect water channel activity. Aquaporins are enriched in zones of fast cell division and expansion, or in areas where water flow or solute flux density would be expected to be high. This includes biotrophic interfaces between plants and parasites, between plants and symbiotic bacteria or fungi, and between germinating pollen and stigma. On a cellular level aquaporin clusters have been identified in some membranes. There is also a possibility that aquaporins in the endoplasmic reticulum may function in symplasmic transport if water can flow from cell to cell via the desmotubules in plasmodesmata. Functional characterization of aquaporins in the native membrane has raised doubt about the conclusiveness of expression patterns alone and need to be conducted in parallel. The challenge will be to elucidate gating on a molecular level and cellular level and to tie those findings into plant water relations on a macroscopic scale where various flow pathways need to be considered.

Plant aquaporins: multifunctional water and solute channels with expanding roles.

The phospholipids that form the membranes of modern cells present a formidable barrier to polar and charged molecules, necessitating complex channels and pumps to permit the exchange of molecules with the external environment. This presents a problem when trying to imagine what a primitive cell would have looked like earlier in the evolution of life. With no transport machinery, how could simple cells take in complex nutrients? A possible answer comes in the form of a model 'protocell' produced by a team at Harvard. Fatty acids and their derivatives are attractive candidates as components of early protocell membranes as they are simple amphiphiles that form bilayer membrane vesicles. A proto-cell equipped with such a membrane is found to be able to take in charged molecules such as nucleotides, while retaining longer genetic polymers made from them inside. The membranes of modern cells are made of phospholipids, which are formidable barriers to the uptake of polar and charged molecules, a challenge to our understanding of the origins of cellular life. Membranes made of simple amphiphiles, such as fatty acids, allow the passage of charged molecules (such as nucleotides), while retaining longer genetic polymers made from them inside such protocells. Primitive cells could thus have acquired complex nutrients from their environment in the absence of any transport machinery. Contemporary phospholipid-based cell membranes are formidable barriers to the uptake of polar and charged molecules ranging from metal ions to complex nutrients. Modern cells therefore require sophisticated protein channels and pumps to mediate the exchange of molecules with their environment. The strong barrier function of membranes has made it difficult to understand the origin of cellular life and has been thought to preclude a heterotrophic lifestyle for primitive cells. Although nucleotides can cross dimyristoyl phosphatidylcholine membranes through defects formed at the gel-to-liquid transition temperature1,2, phospholipid membranes lack the dynamic properties required for membrane growth. Fatty acids and their corresponding alcohols and glycerol monoesters are attractive candidates for the components of protocell membranes because they are simple amphiphiles that form bilayer membrane vesicles3,4,5 that retain encapsulated oligonucleotides3,6 and are capable of growth and division7,8,9. Here we show that such membranes allow the passage of charged molecules such as nucleotides, so that activated nucleotides added to the outside of a model protocell spontaneously cross the membrane and take part in efficient template copying in the protocell interior. The permeability properties of prebiotically plausible membranes suggest that primitive protocells could have acquired complex nutrients from their environment in the absence of any macromolecular transport machinery; that is, they could have been obligate heterotrophs.

Template-directed Synthesis of a Genetic Polymer in a Model Protocell

Several barrier epithelia such as renal collecting duct, urinary bladder, and gastric mucosa maintain high osmotic pH and solute gradients between body compartments and the blood by means of apical membranes of exceptionally low permeabilities. Although the mechanisms underlying these low permeabilities have been only poorly defined, low fluidity of the apical membrane has been postulated. The solubility diffusion model predicts that lower membrane fluidity will reduce permeability by reducing the ability of permeant molecules to diffuse through the lipid bilayer. However, little data compare membrane fluidity with permeability properties, and it is unclear whether fluidity determines permeability to all, or only some substances. We therefore studied the permeabilities of a series of artificial large unilamellar vesicles (LUV) of eight different compositions, exhibiting a range of fluidities encountered in biological membranes. Cholesterol and sphingomyelin content and acyl chain saturation were varied to create a range of fluidities. LUV anisotropy was measured as steady state fluorescence polarization of the lipophilic probe DPH. LUV permeabilities were determined by monitoring concentration-dependent or pH-sensitive quenching of entrapped carboxyfluorescein on a stopped-flow fluorimeter. The relation between DPH anisotropy and permeability to water, urea, acetamide, and NH3 was well fit in each instance by single exponential functions (r > 0.96), with lower fluidity corresponding to lower permeability. By contrast, proton permeability correlated only weakly with fluidity. We conclude that membrane fluidity determines permeability to most nonionic substances and that transmembrane proton flux occurs in a manner distinct from flux of other substances.

https://rupress.org/jgp/article-pdf/106/1/67/1186179/67.pdf

J M Donovan

Papers

The relationship between membrane fluidity and permeabilities to water, solutes, ammonia, and protons.