Small Uncharged Molecules Research Articles

Impact of Polymer Chain Rearrangements in the PA Structure of RO Membranes on Water Permeability and N-Nitrosamine Rejection.

The use of solvents is overall recognized as an efficient method to improve the water permeability of polyamide thin film composite membranes (PA-TFC). The objective of this work was to test the performance of the membranes after exposing them to n-propanol (n-PrOH) to improve the permeability of the membranes while maintaining the rejection factor for small uncharged organic molecules, namely N-nitrosamines (NTRs). After the membranes were exposed to n-PrOH, the water permeability of the UTC73AC membrane increased by 98%, with minimal change in rejection. N-nitrosodiethylamine (NDEA) rejection decreased (3.4%), while N-nitrosodi-n-propylamine (NDPA) and N-nitrosodi-n-butylamine (NDBA) rejection increased by 0.9% and 2.8%, respectively. In contrast, for the BW30LE membrane, water permeability decreased (by 38.7%), while rejection factors increased by 14.5% for NDEA, 6.2% for NDPA, and 15.0% for NDBA. In addition, the morphology of the membrane surface before and after exposure to n-PrOH was analyzed. This result and the pore size distribution (PSD) curves obtained indicate that the rearrangement of polymer chains affects the network or aggregate pores in the PA layer, implying that a change in pore size or a change in pore size distribution could improve the permeability of water molecules, while the rejection factor for NTRs is not significantly affected.

Genome-Wide Identification and Gene Expression Analysis of Sweet Cherry Aquaporins (Prunus avium L.) under Abiotic Stresses.

Aquaporins (AQPs) are integral transmembrane proteins well known as channels involved in the mobilization of water, small uncharged molecules and gases. In this work, the main objective was to carry out a comprehensive study of AQP encoding genes in Prunus avium (cv. Mazzard F12/1) on a genome-wide scale and describe their transcriptional behaviors in organs and in response to different abiotic stresses. A total of 28 non-redundant AQP genes were identified in Prunus spp. Genomes, which were phylogenetically grouped into five subfamilies (seven PIPs, eight NIPs, eight TIPs, three SIPs and two XIPs). Bioinformatic analyses revealed a high synteny and remarkable conservation of structural features among orthologs of different Prunus genomes. Several cis-acting regulatory elements (CREs) related to stress regulation were detected (ARE, WRE3, WUN, STRE, LTR, MBS, DRE, AT-rich and TC-rich). The above could be accounting for the expression variations associated with plant organs and, especially, each abiotic stress analyzed. Gene expressions of different PruavAQPs were shown to be preferentially associated with different stresses. PruavXIP2;1 and PruavXIP1;1 were up-regulated in roots at 6 h and 72 h of hypoxia, and in PruavXIP2;1 a slight induction of expression was also detected in leaves. Drought treatment strongly down-regulated PruavTIP4;1 but only in roots. Salt stress exhibited little or no variation in roots, except for PruavNIP4;1 and PruavNIP7;1, which showed remarkable gene repression and induction, respectively. Interestingly, PruavNIP4;1, the AQP most expressed in cherry roots subjected to cold temperatures, also showed this pattern in roots under high salinity. Similarly, PruavNIP4;2 consistently was up-regulated at 72 h of heat and drought treatments. From our evidence is possible to propose candidate genes for the development of molecular markers for selection processes in breeding programs for rootstocks and/or varieties of cherry.

Aquaporins in Immune Cells and Inflammation: New Targets for Drug Development.

The mammalian immune system senses foreign antigens by mechanisms that involve the interplay of various kinds of immune cells, culminating in inflammation resolution and tissue clearance. The ability of the immune cells to communicate (via chemokines) and to shift shape for migration, phagocytosis or antigen uptake is mainly supported by critical proteins such as aquaporins (AQPs) that regulate water fluid homeostasis and volume changes. AQPs are protein channels that facilitate water and small uncharged molecules’ (such as glycerol or hydrogen peroxide) diffusion through membranes. A number of AQP isoforms were found upregulated in inflammatory conditions and are considered essential for the migration and survival of immune cells. The present review updates information on AQPs’ involvement in immunity and inflammatory processes, highlighting their role as crucial players and promising targets for drug discovery.

Exploiting σ-hole interaction to design small uncharged ligand molecules to stabilize G-quadruplex-DNA: a computational study.

Small neutral seleno molecules were designed to stabilize G-quadruplex-DNA to accelerate the death of cancer cells. A new approach was considered in this study to design new ligands to stabilize G-quadruplex-DNA using the non-covalent σ-hole interaction. The systematic study has been performed with ligands in the absence and presence of σ-hole interaction to stabilize G-tetrad. Fluorine-substituted seleno ligands interact with the bases of G-quadruplex strongly with the σ-holes present in the ligands. The binding of the fluorinated ligands FSeCF2SeCF2 SeF2(4) (~75.0kcal/mol) is stronger than that of BRACO-19 (~70.0kcal/mol) calculated at the same level of theory with G-tetrad. It is reported that BRACO-19 is employed to inhibit the enzymatic activity of telomerase. The calculated results also reveal the importance of optimum chain length of ligands to achieve a better binding ability with G-tetrad. The MESP calculations and AIM analysis corroborate the trends of binding of these ligand molecules with G-quadruplexes. The small neutral molecules possess considerable advantage to pass through the lipid bilayer of the cell membrane by passive transport and are of choice in biological research and for clinical trials. Graphical AbtractSmall neutral ligands with σ-holes can stabilize G-quadruplex to inhibit enzymatic activity of telomerase.

Aquaporins and placenta.

Water is the major component of cells and tissues. The fetal body consists of about 70-90% water and its fluid balance is dependent on the mother. In fact, abortion, premature birth, amniotic fluid volume abnormality, malformation and fetal growth restrictions might result when the homeostasis of the maternal-fetal fluid exchange is disrupted. Thus, maternal-fetal fluid balance is critical during pregnancy. In this sense, several mechanisms, including aquaporins (AQPs) have been reported to play important roles in maternal-fetal fluid balance. AQPs are small membrane proteins (about 30kDa), present in different organs, that increase the permeability of water, as well as other small uncharged molecules to be transported across the bilayer cell membranes. Several aquaporins are expressed in placenta, and aquaporins play key roles in the placental function. Even though aquaporins have a proven crucial role in water homeostasis, the physiological and pathological importance of aquaporins as glycerol channels is not fully understood. This review focuses on advances in our knowledge of the roles of aquaporins in placental cells, particularly the roles of AQP3 and AQP9 in placental metabolism and points to the pathophysiological importance of glycerol channels in placenta, as well as the signal transduction pathways activated by them. Moreover, the regulation of aquaporins expression by different placental hormones, such as leptin and the mechanisms involved will be discussed.

Halogen‐Mediated Membrane Transport: An Efficient Strategy for the Enhancement of Cellular Uptake of Synthetic Molecules

The poor uptake of fluorescent probes and therapeutics by mammalian cells is a major concern in biological applications ranging from fluorescence imaging to drug delivery in living cells. Although gaseous molecules such as oxygen and carbon dioxide, hydrophobic substances such as benzene, and small polar but uncharged molecules such as water and ethanol can cross the cell plasma membrane by simple passive diffusion, many synthetic as well as biological molecules require specific membrane transporters and channel proteins that control the traffic of these molecules into and out of the cell. This work reports that the introduction of halogen atoms into a series of fluorescent molecules remarkably enhances their cellular uptake, and that their transport can be increased to more than 95 % by introducing two iodine atoms at appropriate positions. The nature of the fluorophore does not play a major role in the cellular uptake when iodine atoms are present in the molecules, as compounds bearing naphthalimide, coumarin, BODIPY, and pyrene moieties show similar uptakes. Interestingly, the introduction of a maleimide-based fluorophore bearing two hydroxyethylthio moieties allows the molecules to cross the plasma and nuclear membranes, and the presence of iodine atoms further enhances the transport across both membranes. Overall, this study provides a general strategy for enhancing the uptake of organic molecules by mammalian cells.

Quantification of the Intracellular Life Time of Water Molecules to Measure Transport Rates of Human Aquaglyceroporins

Orthodox aquaporins are transmembrane channel proteins that facilitate rapid diffusion of water, while aquaglyceroporins facilitate the diffusion of small uncharged molecules such as glycerol and arsenic trioxide. Aquaglyceroporins play important roles in human physiology, in particular for glycerol metabolism and arsenic detoxification. We have developed a unique system applying the strain of the yeast Pichia pastoris, where the endogenous aquaporins/aquaglyceroporins have been removed and human aquaglyceroporins AQP3, AQP7, and AQP9 are recombinantly expressed enabling comparative permeability measurements between the expressed proteins. Using a newly established Nuclear Magnetic Resonance approach based on measurement of the intracellular life time of water, we propose that human aquaglyceroporins are poor facilitators of water and that the water transport efficiency is similar to that of passive diffusion across native cell membranes. This is distinctly different from glycerol and arsenic trioxide, where high glycerol transport efficiency was recorded.

Putative new groups of invertebrate water channels based on the snail Helix pomatia L. (Helicidae) MIP protein identification and phylogenetic analysis

Water channel proteins, classified as a family of Membrane Intrinsic Proteins (MIPs) superfamily, enable rapid movement of water and small uncharged molecules through biological membranes. Although water channel proteins are required in several important processes characteristic for the animals, such as osmoregulation, mucus secretion, or defense against desiccation, molluscs, until now, have been very poorly explored in this aspect. Therefore, we decided to study MIPs in Helix pomatia L. applied as a model in studies on terrestrial snail physiology.Our studies consisted in: the snail organ transcriptome sequencing and consecutive bioinformatic analysis of the predicted protein, estimation of the encoding transcript expression (qPCR), investigation of the predicted protein function in the yeast Saccharomyces cerevisiae cells, and the phylogenetic analysis.We identified six water channel proteins, named HpAQP1 to HpAQP6. All of them were proven to transport water, two of them (HpAQP3 and HpAQP4) were also shown to be able to transport glycerol, and other two (HpAQP5 and HpAQP6) to transport H2O2. Phylogenetic analysis indicated that the proteins either fell into aquaporins (HpAQP1, HpAQP2 and HpAQP5) or formed new groups of invertebrate water channel proteins, not described until now, that we suggest to term malacoglyceroporins (HpAQP3 and HpAQP4) and malacoaquaporins (HpAQP6). Thus, the classification of animal water channels based on the vertebrate proteins and including aquaporin, aquaglyceroporin, S-aquaporin and AQP8-type grades does not reflect diversity of these proteins in invertebrates.The obtained results provide important data concerning diversity of water channel protein repertoire in aquatic and terrestrial invertebrates and should also contribute to the improvement of animal water channel classification system.

Molecular Effects of Concentrated Solutes on Protein Hydration, Dynamics, and Electrostatics

Most studies of protein structure and function are performed in dilute conditions, but proteins typically experience high solute concentrations in their physiological scenarios and biotechnological applications. High solute concentrations have well-known effects on coarse protein traits like stability, diffusion, and shape, but likely also perturb other traits through finer effects pertinent at the residue and atomic levels. Here, NMR and molecular dynamics investigations on ubiquitin disclose variable interactions with concentrated solutes that lead to localized perturbations of the protein’s surface, hydration, electrostatics, and dynamics, all dependent on solute size and chemical properties. Most strikingly, small polar uncharged molecules are sticky on the protein surface, whereas charged small molecules are not, but the latter still perturb the internal protein electrostatics as they diffuse nearby. Meanwhile, interactions with macromolecular crowders are favored mainly through hydrophobic, but not through polar, surface patches. All the tested small solutes strongly slow down water exchange at the protein surface, whereas macromolecular crowders do not exert such strong perturbation. Finally, molecular dynamics simulations predict that unspecific interactions slow down microsecond- to millisecond-timescale protein dynamics despite having only mild effects on pico- to nanosecond fluctuations as corroborated by NMR. We discuss our results in the light of recent advances in understanding proteins inside living cells, focusing on the physical chemistry of quinary structure and cellular organization, and we reinforce the idea that proteins should be studied in native-like media to achieve a faithful description of their function.

Role of Electroosmosis in the Permeation of Neutral Molecules: CymA and Cyclodextrin as an Example

To quantify the flow of small uncharged molecules into and across nanopores, one often uses ion currents. The respective ion-current fluctuations caused by the presence of the analyte make it possible to draw some conclusions about the direction and magnitude of the analyte flow. However, often this flow appears to be asymmetric with respect to the applied voltage. As a possible reason for this asymmetry, we identified the electroosmotic flow (EOF), which is the water transport associated with ions driven by the external transmembrane voltage. As an example, we quantify the contribution of the EOF through a nanopore by investigating the permeation of α-cyclodextrin through CymA, a cyclodextrin-specific channel from Klebsiella oxytoca. To understand the results from electrophysiology on a molecular level, all-atom molecular dynamics simulations are used to detail the effect of the EOF on substrate entry to and exit from a CymA channel in which the N-terminus has been deleted. The combined experimental and computational results strongly suggest that one needs to account for the significant contribution of the EOF when analyzing the penetration of cyclodextrins through the CymA pore. This example study at the same time points to the more general finding that the EOF needs to be considered in translocation studies of neutral molecules and, at least in many cases, should be able to help in discriminating between translocation and binding events.

Membrane inlet mass spectrometry reveals that Ceriporiopsis subvermispora bicupin oxalate oxidase is inhibited by nitric oxide

Membrane inlet mass spectrometry (MIMS) uses a semipermeable membrane as an inlet to a mass spectrometer for the measurement of the concentration of small uncharged molecules in solution. We report the use of MIMS to characterize the catalytic properties of oxalate oxidase (E.C. 1.2.3.4) from Ceriporiopsis subvermispora (CsOxOx). Oxalate oxidase is a manganese dependent enzyme that catalyzes the oxygen-dependent oxidation of oxalate to carbon dioxide in a reaction that is coupled with the formation of hydrogen peroxide. CsOxOx is the first bicupin enzyme identified that catalyzes this reaction. The MIMS method of measuring OxOx activity involves continuous, real-time direct detection of oxygen consumption and carbon dioxide production from the ion currents of their respective mass peaks. 13C2-oxalate was used to allow for accurate detection of 13CO2 (m/z 45) despite the presence of adventitious 12CO2. Steady-state kinetic constants determined by MIMS are comparable to those obtained by a continuous spectrophotometric assay in which H2O2 production is coupled to the horseradish peroxidase catalyzed oxidation of 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid). Furthermore, we used MIMS to determine that NO inhibits the activity of the CsOxOx with a KI of 0.58±0.06μM.

Aquaporins: for more than water at the plant–fungus interface?

Aquaporins are abundant membrane proteins that channel water, but also small uncharged molecules, across the membranes of plants, animals and microbes (Maurel et al., 2008). In this issue of New Phytologist, Dietz et al. (pp. 927–940) report on a thorough molecular and functional analysis of the whole aquaporin family of an ectomycorrhizal (ECM) fungus, the basidiomycete Laccaria bicolor. This study is novel as it describes, from an evolutionary perspective, the diversity of aquaporins in a fungus, which, with the exception of the yeast Saccharomyces cerevisiae, had not been extensively studied at a genome scale in these organisms. This study is also of special importance as it addresses the role of aquaporins in both nutrient and water exchanges during symbiosis between plants and fungi, with a potentially great impact on water and nutrient economy in forest ecosystems. In brief, Dietz et al. screened the full genomic sequence of Laccaria bicolor to identify seven open reading frames (ORFs) encoding aquaporin homologues. They used heterologous expression in yeast and Xenopus oocytes to characterize the ability of these homologues to transport water but also various solutes such as glycerol, urea andammonium (NH4+)/ammonia (NH3). It is of note that nitrogen (N) compounds, some of which are putative aquaporin substrates, are, besides water, crucial during ECM symbiosis. Similar to observations of aquaporins in other organisms, various selectivity profiles were identified for the L. bicolor aquaporins; some isoforms specifically transported water whereas others were also permeable to urea, glycerol and/or NH3.

Carbon nanotubes-based chemiresistive immunosensor for small molecules: Detection of nitroaromatic explosives

In recent years, there has been a growing focus on use of one-dimensional (1-D) nanostructures, such as carbon nanotubes and nanowires, as transducer elements for label-free chemiresistive/field-effect transistor biosensors as they provide label-free and high sensitivity detection. While research to-date has elucidated the power of carbon nanotubes- and other 1-D nanostructure-based field effect transistors immunosensors for large charged macromolecules such as proteins and viruses, their application to small uncharged or charged molecules has not been demonstrated. In this paper we report a single-walled carbon nanotubes (SWNTs)-based chemiresistive immunosensor for label-free, rapid, sensitive and selective detection of 2,4,6-trinitrotoluene (TNT), a small molecule. The newly developed immunosensor employed a displacement mode/format in which SWNTs network forming conduction channel of the sensor was first modified with trinitrophenyl (TNP), an analog of TNT, and then ligated with the anti-TNP single chain antibody. Upon exposure to TNT or its derivatives the bound antibodies were displaced producing a large change, several folds higher than the noise, in the resistance/conductance of SWNTs giving excellent limit of detection, sensitivity and selectivity. The sensor detected between 0.5 ppb and 5000 ppb TNT with good selectivity to other nitroaromatic explosives and demonstrated good accuracy for monitoring TNT in untreated environmental water matrix. We believe this new displacement format can be easily generalized to other one-dimensional nanostructure-based chemiresistive immuno/affinity-sensors for detecting small and/or uncharged molecules of interest in environmental monitoring and health care.

Exploring Transmembrane Diffusion Pathways With Molecular Dynamics

Transmembrane exchange of materials is a fundamental process in biology. Molecular dynamics provides a powerful method to investigate in great detail various aspects of the phenomenon, particularly the permeation of small uncharged molecules, which continues to pose a challenge to experimental studies. We will discuss some of the recent simulation studies investigating the role of lipid-mediated and protein-mediated mechanisms in permeation of water and gas molecules across the membrane.

A Current View of the Mammalian Aquaglyceroporins

The discovery of aquaporin water channels by Agre and coworkers answered a long-standing biophysical question of how the majority of water crosses biological membranes. The identification and study of aquaporins have provided insight, at the molecular level, into the fundamental physiology of water balance regulation and the pathophysiology of water balance disorders. In addition to the originally identified classical aquaporins, a second class of aquaporins has been identified. Aquaporins in this latter class, the so-called aquaglyceroporins, transport small uncharged molecules such as glycerol and urea as well as water. Aquaglyceroporins have a wide tissue distribution, and emerging data suggest that several of them may play previously unappreciated physiological or pathophysiological roles. Analyses of transgenic mice have revealed potential roles of aquaglyceroporins in skin elasticity, gastrointestinal function and metabolism, and metabolic diseases such as diabetes mellitus. This review comprehensively discusses the recent discoveries in the field of aquaglyceroporins, alongside a brief overview of the so-called unorthodox aquaporins.

The aquaporins

Aquaporins are intrinsic membrane proteins found in all organisms, from archaea to mammals. They selectively allow water or other small uncharged molecules to pass along the osmotic gradient.

Crossing the Bilayer

A ll cells, whether bacterial, plant, or animal, are enclosed by membranes, the basic components of which are lipid bilayers. The cell membrane ultimately acts as the defining principle of what constitutes a cell and what constitutes the rest of the world. Lipid bilayers are semipermeable: Small uncharged molecules can pass more or less freely from one side of the membrane to the other, but for charged species or macromolecules, such as proteins and DNA, the lipid bilayer is a major obstacle to diffusion. However, in real life, cells need to be able to transport proteins, DNA, and ions into and out of the cell, across the lipid bilayer. In this special issue, we look at the mechanisms used by cells to allow proteins, DNA, and ions to directly traverse a biological membrane. Wickner and Schekman (p. 1452) describe how proteins can cross, or become integrated into, specific membranes. The endoplasmic reticulum membrane in eukaryotes and the plasma membrane in bacteria contain a proteinaceous pore—the translocon—that specifically promotes the translocation and integration of a multitude of signal-sequence-bearing membrane and secretory proteins into and through the membranes. Beyond these canonical systems that use the translocon, the authors mention translocation machineries and targeting strategies involved in import into different organelles, such as the mitochondria, chloroplasts, and peroxisomes, within cells and across kingdoms. Proteins are not the only macromolecules transferred across the membrane. Chen et al. (p. 1456) describe how bacteria allow DNA to traverse their membranes during the processes of conjugation and transformation and compare and contrast the molecular machineries involved in targeting and transport. Regulating the internal composition of the cytosol is a key process in maintaining the chemistry of life, and two of the most fundamental components are the concentration and intracellular/extracellular balance of a variety of biologically important ions, including sodium, potassium, calcium, and chloride. Gouaux and MacKinnon (p. 1461) describe how ions get across membranes via transmembrane pumps and channels and explain the chemical and structural constraints involved. They describe the importance of gating within these structures, which allows for the very high specificity of transport observed and for the establishment and maintenance of important electrochemical gradients across cell membranes. Science 's Signal Transduction Knowledge Environment (STKE, [stke.sciencemag.org][1]) addresses how information is transmitted across cell membranes to contribute to cell signaling processes. When ions and proteins cross cell membranes, they may trigger intracellular signaling cascades. Hisatsune and Mikoshiba describe how in mammalian cells, the inositol phosphate receptor (the IP3 receptor) and the calcium sensor (STIM) redistribute and cluster in response to changes in intracellular calcium and mediate calcium influx into the endoplasmic reticulum to refill intracellular calcium stores. Joliot describes how a class of cell-penetrating peptides can allow cells to communicate with one another, and Onfelt et al. describe how nanotubular membrane connections mediate intercellular communication. The Teaching Resource by Felsenfeld provides lecture materials describing how integrins transmit information from the extracellular matrix to the cytoskeleton. How cells generate and maintain their internal structures and integrity depends in large part on the effectiveness of the membrane in keeping the inside in and the outside out. The mechanisms used in the transfer of ions and macromolecules across the cell membrane can thus be considered one of the defining principles of life, and our understanding of these processes is fundamental to our understanding of all other aspects of cellular and organismal physiology. [1]: http://stke.sciencemag.org/

Modulation of GABAergic Transmission by Activity via Postsynaptic Ca2+-Dependent Regulation of KCC2 Function

Activity-induced modification of GABAergic transmission contributes to the plasticity of neural circuits. In the present work we found that prolonged postsynaptic spiking of hippocampal neurons led to a shift in the reversal potential of GABA-induced Cl- currents (E(Cl)) toward positive levels in a duration- and frequency-dependent manner. This effect was abolished by blocking cytosolic Ca2+ elevation and mimicked by releasing Ca2+ from internal stores. Activity- and Ca2+-induced E(Cl) shifts were larger in mature neurons, which express the K-Cl cotransporter KCC2 at high levels, and inhibition of KCC2 occluded the shifts. Overexpression of KCC2 in young cultured neurons, which express lower levels of KCC2 and have a more positive E(Cl), resulted in hyperpolarized E(Cl) similar to that of mature cells. Importantly, these young KCC2-expressing neurons became responsive to neuronal spiking and Ca2+ elevation by showing positive E(Cl) shifts. Thus, repetitive postsynaptic spiking reduces the inhibitory action of GABA through a Ca2+-dependent downregulation of KCC2 function.

Estimating the plasma membrane permeability of Taxus x media cells with the spin probe TEMPOL by EPR

Production of paclitaxel and other taxanes by cell suspension cultures of Taxus species is an alternative to their extraction from yew tree bark or shoots. Little is known about the mechanism of taxane transport in Taxus cells, in either cell cultures or plants. In our study we measured the influence of jasmonic acid (JA), an elicitor of taxane production, on plasma membrane permeability in cell suspensions of Taxus x media. TEMPOL spin probe was added to the suspension of cells grown in the presence of JA and to control cells. Reduction of TEMPOL in the cell interior was measured by electron paramagnetic resonance (EPR). Plasma membrane permeability was derived from the kinetics of reduction. JA was shown to decrease the membrane permeability of Taxus x media cells. The observed inhibition of the spin probe permeability by JA can be ascribed to modification of the membrane structure and lipid composition or, at least, to changes in the lipid lateral domain structures, thus affecting the transport of TEMPOL. The changed permeability to small uncharged molecules could influence the primary cellular metabolism and possibly also the transport of taxanes.

The glycerol facilitator GlpF, its aquaporin family of channels, and their selectivity

Publisher Summary The “glycerol facilitator” GlpF, a highly selective transmembrane channel that conducts glycerol and certain other small uncharged organic molecules, was the first member of the large aquaporin family (AQP family) to be identified at the genetic level and functionally characterized. Once glycerol is passaged inside the cell, it is rapidly phosphorylated by glycerol kinase to produce glycerol 3-phosphate that proceeds by dehydrogenation to dihydroxyacetone phosphate (DHAP) or to phospholipid synthesis where glycerol provides the basis for attachment of fatty acid chains and phophatidyl head groups in ∼2/3 of cellular phospholipids. The glp regulon is inducible by glycerol 3-phosphate which provides an advantage for growth of Escherichia coli on glycerol. The glpF gene is the first gene on the glp operon that also contains the gene for glycerol kinase.