Arthur Hunt | University of Kentucky (original) (raw)
Papers by Arthur Hunt
Frontiers in Plant Science, Jan 21, 2024
The Metabolism, Structure, and Function of Plant Lipids, 1987
Lipoxygenases (LOXs) represent a group of polyunsaturated fatty acid oxidases that are apparently... more Lipoxygenases (LOXs) represent a group of polyunsaturated fatty acid oxidases that are apparently ubiquitious in eukaryotic organisms. There are a growing number of reports of the presence of this enzyme in various human and other animal tissues. The list of plants containing active LOXs also continues to expand.
Here we present major findings of a national survey of faculty teaching genomics CUREs conducted ... more Here we present major findings of a national survey of faculty teaching genomics CUREs conducted by the GEA, and our work on presenting relevant resources on the QUBES web portal.
Wiley Interdisciplinary Reviews - Rna, Nov 7, 2019
Journal of Biological Chemistry, Dec 1, 1981
The reconstitution of binding protein-dependent glutamine transport in isolated membrane vesicles... more The reconstitution of binding protein-dependent glutamine transport in isolated membrane vesicles from Escherichia coli is described. The reconstituted glutamine transport is shown to be energy-dependent and does not involve the metabolism of glutamine or the trapping of liganded binding protein with the vesicles. The preparation of vesicles capable of transporting glutamine in a binding protein-dependent manner required the incorporation of NAD into vesicles and the use of a binding protein point mutant as the source of the vesicles.
Biochemistry, Feb 15, 1983
We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide... more We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide (NBS) to modify respectively the sole histidine and tryptophan residues and examined the effect of these modifications on the ability of the binding protein to bind glutamine as well as the ability to restore glutamine transport in membrane vesicles of Escherichia coli. Under the conditions used, both DEPC and NBS markedly inhibited the ability to restore glutamine transport in vesicles without any significant effect on glutamine binding. Moreover, saturating quantities of glutamine had no protective effect on the inactivation of the binding protein by DEPC or NBS. Fluorometric measurement and amino acid analysis indicate that the inactivation of the binding protein in restoring vesicle transport by NBS can be attributed to the oxidation of a single tryptophan residue. Similar analysis and the inability of hydroxylamine to reverse the effect of DEPC indicate that the effects of DEPC can probably be attributed to alterations of the sole histidine and/or one or more lysine residues of the binding protein. We conclude that the glutamine binding protein possesses at least two largely nonoverlapping functional domains, one responsible for glutamine binding and the other for the interaction with the other components of the glutamine transport system.
Research Square (Research Square), Aug 19, 2019
The Model Legume Medicago truncatula, Dec 13, 2019
Methods in Enzymology, 1986
Publisher Summary This chapter examines how several bacterial active transport systems require, i... more Publisher Summary This chapter examines how several bacterial active transport systems require, in addition to one or more membrane-bound components, periplasmic binding proteins. Genetic studies, experiments with intact cells, and investigations using spheroplasts have all established the unique role that these proteins play in the transport process. The chapter describes methods for preparing isolated membrane vesicles capable of binding protein-dependent glutamine transport, and for assaying glutamine transport in such vesicles. Two different kinds of mutants are essential for the study of binding protein-dependent transport in isolated membrane vesicles. First, a strain that overproduces the various components of the system under study is needed, both to provide a ready source of the substrate binding protein, and to serve as a parent for the second type of strain. Second, a derivative of the overproducing strain that has a point mutation in the binding protein gene is needed to serve as a source of membrane vesicles. Glutamine-binding protein is purified from a strain that overproduces the various components of the glutamine transport system.
<p>The occurrences of 5 nt motifs was determined using SignalSleuth2 [<a href="http... more <p>The occurrences of 5 nt motifs was determined using SignalSleuth2 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146107#pone.0146107.ref051" target="_blank">51</a>]. The relative position of the motif is given on the x-axis, with the poly(A) site being set as “0”; in these plots, regions extending from 80 nts 5’ (upstream) to 80 nts 3’ (downstream” from the poly(A) sites are shown. The numerical count of each motif is given on the y-axis. The plot shows the distributions of the 50 most-abundant motifs, which are listed in the legend embedded on the right. The number of sites used for this analysis was 39,415. (A) Profiles of the 10 most abundant motifs. The individual motifs are noted in the caption in the upper right corner. (B) Profiles of the next 40 most abundant motifs. The individual motifs are noted in the caption on the right. Note that the y-axis scale for panel B is 10% of that for panel A.</p
Biochemistry, 1983
We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide... more We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide (NBS) to modify respectively the sole histidine and tryptophan residues and examined the effect of these modifications on the ability of the binding protein to bind glutamine as well as the ability to restore glutamine transport in membrane vesicles of Escherichia coli. Under the conditions used, both DEPC and NBS markedly inhibited the ability to restore glutamine transport in vesicles without any significant effect on glutamine binding. Moreover, saturating quantities of glutamine had no protective effect on the inactivation of the binding protein by DEPC or NBS. Fluorometric measurement and amino acid analysis indicate that the inactivation of the binding protein in restoring vesicle transport by NBS can be attributed to the oxidation of a single tryptophan residue. Similar analysis and the inability of hydroxylamine to reverse the effect of DEPC indicate that the effects of DEPC can probably be attributed to alterations of the sole histidine and/or one or more lysine residues of the binding protein. We conclude that the glutamine binding protein possesses at least two largely nonoverlapping functional domains, one responsible for glutamine binding and the other for the interaction with the other components of the glutamine transport system.
Methods in molecular biology, Nov 13, 2014
Second generation DNA sequencing technologies have been a great boon for the study of mRNA polyad... more Second generation DNA sequencing technologies have been a great boon for the study of mRNA polyadenylation. The experimental determination of large numbers of polyadenylation sites using high-throughput sequencing strategies has provided the necessary platform for deeper understanding the regulation of gene expression in eukaryotes. For generating large sets of data to map poly-A sites, specialized sample preparations that target the junction of 3&amp;amp;amp;#39;-UTR and the poly(A) tail are usually employed. Here, we describe three different protocols that are effectively used for global determinations of poly(A) site choice in plants.
Frontiers in Plant Science, 2012
Nucleic Acids Research, 1991
Plant Physiology, Oct 1, 1997
Journal of Virology, 1991
Frontiers in Plant Science, Jan 21, 2024
The Metabolism, Structure, and Function of Plant Lipids, 1987
Lipoxygenases (LOXs) represent a group of polyunsaturated fatty acid oxidases that are apparently... more Lipoxygenases (LOXs) represent a group of polyunsaturated fatty acid oxidases that are apparently ubiquitious in eukaryotic organisms. There are a growing number of reports of the presence of this enzyme in various human and other animal tissues. The list of plants containing active LOXs also continues to expand.
Here we present major findings of a national survey of faculty teaching genomics CUREs conducted ... more Here we present major findings of a national survey of faculty teaching genomics CUREs conducted by the GEA, and our work on presenting relevant resources on the QUBES web portal.
Wiley Interdisciplinary Reviews - Rna, Nov 7, 2019
Journal of Biological Chemistry, Dec 1, 1981
The reconstitution of binding protein-dependent glutamine transport in isolated membrane vesicles... more The reconstitution of binding protein-dependent glutamine transport in isolated membrane vesicles from Escherichia coli is described. The reconstituted glutamine transport is shown to be energy-dependent and does not involve the metabolism of glutamine or the trapping of liganded binding protein with the vesicles. The preparation of vesicles capable of transporting glutamine in a binding protein-dependent manner required the incorporation of NAD into vesicles and the use of a binding protein point mutant as the source of the vesicles.
Biochemistry, Feb 15, 1983
We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide... more We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide (NBS) to modify respectively the sole histidine and tryptophan residues and examined the effect of these modifications on the ability of the binding protein to bind glutamine as well as the ability to restore glutamine transport in membrane vesicles of Escherichia coli. Under the conditions used, both DEPC and NBS markedly inhibited the ability to restore glutamine transport in vesicles without any significant effect on glutamine binding. Moreover, saturating quantities of glutamine had no protective effect on the inactivation of the binding protein by DEPC or NBS. Fluorometric measurement and amino acid analysis indicate that the inactivation of the binding protein in restoring vesicle transport by NBS can be attributed to the oxidation of a single tryptophan residue. Similar analysis and the inability of hydroxylamine to reverse the effect of DEPC indicate that the effects of DEPC can probably be attributed to alterations of the sole histidine and/or one or more lysine residues of the binding protein. We conclude that the glutamine binding protein possesses at least two largely nonoverlapping functional domains, one responsible for glutamine binding and the other for the interaction with the other components of the glutamine transport system.
Research Square (Research Square), Aug 19, 2019
The Model Legume Medicago truncatula, Dec 13, 2019
Methods in Enzymology, 1986
Publisher Summary This chapter examines how several bacterial active transport systems require, i... more Publisher Summary This chapter examines how several bacterial active transport systems require, in addition to one or more membrane-bound components, periplasmic binding proteins. Genetic studies, experiments with intact cells, and investigations using spheroplasts have all established the unique role that these proteins play in the transport process. The chapter describes methods for preparing isolated membrane vesicles capable of binding protein-dependent glutamine transport, and for assaying glutamine transport in such vesicles. Two different kinds of mutants are essential for the study of binding protein-dependent transport in isolated membrane vesicles. First, a strain that overproduces the various components of the system under study is needed, both to provide a ready source of the substrate binding protein, and to serve as a parent for the second type of strain. Second, a derivative of the overproducing strain that has a point mutation in the binding protein gene is needed to serve as a source of membrane vesicles. Glutamine-binding protein is purified from a strain that overproduces the various components of the glutamine transport system.
<p>The occurrences of 5 nt motifs was determined using SignalSleuth2 [<a href="http... more <p>The occurrences of 5 nt motifs was determined using SignalSleuth2 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146107#pone.0146107.ref051" target="_blank">51</a>]. The relative position of the motif is given on the x-axis, with the poly(A) site being set as “0”; in these plots, regions extending from 80 nts 5’ (upstream) to 80 nts 3’ (downstream” from the poly(A) sites are shown. The numerical count of each motif is given on the y-axis. The plot shows the distributions of the 50 most-abundant motifs, which are listed in the legend embedded on the right. The number of sites used for this analysis was 39,415. (A) Profiles of the 10 most abundant motifs. The individual motifs are noted in the caption in the upper right corner. (B) Profiles of the next 40 most abundant motifs. The individual motifs are noted in the caption on the right. Note that the y-axis scale for panel B is 10% of that for panel A.</p
Biochemistry, 1983
We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide... more We treated the glutamine binding protein with diethyl pyrocarbonate (DEPC) and N-bromosuccinimide (NBS) to modify respectively the sole histidine and tryptophan residues and examined the effect of these modifications on the ability of the binding protein to bind glutamine as well as the ability to restore glutamine transport in membrane vesicles of Escherichia coli. Under the conditions used, both DEPC and NBS markedly inhibited the ability to restore glutamine transport in vesicles without any significant effect on glutamine binding. Moreover, saturating quantities of glutamine had no protective effect on the inactivation of the binding protein by DEPC or NBS. Fluorometric measurement and amino acid analysis indicate that the inactivation of the binding protein in restoring vesicle transport by NBS can be attributed to the oxidation of a single tryptophan residue. Similar analysis and the inability of hydroxylamine to reverse the effect of DEPC indicate that the effects of DEPC can probably be attributed to alterations of the sole histidine and/or one or more lysine residues of the binding protein. We conclude that the glutamine binding protein possesses at least two largely nonoverlapping functional domains, one responsible for glutamine binding and the other for the interaction with the other components of the glutamine transport system.
Methods in molecular biology, Nov 13, 2014
Second generation DNA sequencing technologies have been a great boon for the study of mRNA polyad... more Second generation DNA sequencing technologies have been a great boon for the study of mRNA polyadenylation. The experimental determination of large numbers of polyadenylation sites using high-throughput sequencing strategies has provided the necessary platform for deeper understanding the regulation of gene expression in eukaryotes. For generating large sets of data to map poly-A sites, specialized sample preparations that target the junction of 3&amp;amp;amp;#39;-UTR and the poly(A) tail are usually employed. Here, we describe three different protocols that are effectively used for global determinations of poly(A) site choice in plants.
Frontiers in Plant Science, 2012
Nucleic Acids Research, 1991
Plant Physiology, Oct 1, 1997
Journal of Virology, 1991