Journal of magnetic resonance (San Diego, Calif. : 1997), 2007
The inductance of a radiofrequency coil determines its compatibility with a given NMR probe circu... more The inductance of a radiofrequency coil determines its compatibility with a given NMR probe circuit. However, calculation (or estimation) of inductance for radiofrequency coils of dimensions suitable for use in an NMR probe is not trivial, particularly for flat-coils. A comparison of a number of formulae for calculation of inductance is presented through the use of a straightforward inductance measurement circuit. This technique relies upon instrumentation available in many NMR laboratories rather than upon more expensive and specialized instrumentation often utilized in the literature. Inductance estimation methods are suggested and validated for both flat-coils and solenoids. These have proven very useful for fabrication of a number of new coils in our laboratory for use in static solid-state NMR probes operating at (1)H frequencies of 300 and 600MHz. Solenoidal coils with very similar measured and estimated inductances having inner diameters from 1 to 5mm are directly compared as...
The reovirus p10 fusion-associated small transmembrane (FAST) proteins are the smallest known mem... more The reovirus p10 fusion-associated small transmembrane (FAST) proteins are the smallest known membrane fusion proteins, and evolved specifically to mediate cell-cell, rather than virus-cell, membrane fusion. The 36-40-residue ectodomains of avian reovirus (ARV) and Nelson Bay reovirus (NBV) p10 contain an essential intramolecular disulfide bond required for both cell-cell fusion and lipid mixing between liposomes. To more clearly define the functional, biochemical and biophysical features of this novel fusion peptide, synthetic peptides representing the p10 ectodomains of ARV and NBV were analyzed by solution-state NMR spectroscopy, circular dichroism spectroscopy, fluorescence spectroscopy-based hydrophobicity analysis, and liposome binding and fusion assays. Results indicate that disulfide bond formation promotes exposure of hydrophobic residues, as indicated by bis-ANS binding and time-dependent peptide aggregation under aqueous conditions, implying the disulfide bond creates a s...
A holder allowing rotation of a flat coil relative to the static magnetic field (B0) of a nuclear... more A holder allowing rotation of a flat coil relative to the static magnetic field (B0) of a nuclear magnetic resonance (NMR) spectrometer over the range of orientations from parallel to perpendicular has been built and incorporated into a two-channel wide line static solid-state NMR probe. Radiofrequency induction by the coil is perpendicular to B0 for all rotational angles. Such a
Spider silk is one of nature&... more Spider silk is one of nature's most remarkable biomaterials due to extraordinary strength and toughness not found in today's synthetic materials. Of the seven types of silk, wrapping silk (AcSp1) is the most extensible of the types of silks and has no sequence similarity to the other types. Here we report the chemical shifts for the AcSp1 199 amino acid protein repeat unit and its anticipated secondary structure based on secondary chemical shifts.
+ /H + exchanger) is a ubiquitously ex- pressed integral membrane protein that regulates intracel... more + /H + exchanger) is a ubiquitously ex- pressed integral membrane protein that regulates intracellular pH by removing a proton in exchange for an extracellular sodium ion. Of the nine known isoforms of the mammalian NHEs, the firstisoformdiscovered(NHE1)isthemostthoroughlycharacter- ized. NHE1 is involved in numerous physiological processes in mammals, including regulation of intracellular pH, cell-volume control, cytoskeletal organization, heart disease and cancer. NHE comprises two domains: an N-terminal membrane domain that functions to transport ions, and a C-terminal cytoplasmic regul- atory domain that regulates the activity and mediates cytoskeletal interactions.AlthoughtheexactmechanismoftransportbyNHE1 remainselusive,recentstudieshaveidentifiedaminoacidresidues that are important for NHE function. In addition, progress has been made regarding the elucidation of the structure of NHEs. Specifically, the structure of a single TM (transmembrane) seg- ment from NHE1 has been sol...
Rhomboid proteases are integral membrane serine proteases that catalyze peptide bond hydrolysis i... more Rhomboid proteases are integral membrane serine proteases that catalyze peptide bond hydrolysis in biological membranes. Little is currently known about the interaction of enzyme and substrate. Coarse-grained molecular dynamics simulations in hydrated lipid bilayers are employed herein to study the interaction of the E. coli rhomboid protease GlpG (ecGlpG) with the transmembrane domain (TMD) of the substrate Spitz. Spitz does not associate with ecGlpG exclusively at the putative substrate gate near TMD 5. Instead, there are six prominent and stable interaction sites, including one between TMDs 1 and 3, with the closest enzyme-substrate proximity occurring at the ends of helical TMDs or in loops. Bilayer thinning is observed proximal to ecGlpG, but there is no evidence of additional thinning of the bilayer upon interaction with substrate. We suggest that the initial interaction between enzyme and substrate, or substrate capture event, is not limited to a single site on the enzyme, and may be driven by juxtamembrane electrostatic interactions. The findings are of additional interest because catalytically inactive rhomboids (iRhoms) are now known to interact with the substrates of their catalytically active counterparts and to antagonize the enzyme-driven pathways.
Journal of magnetic resonance (San Diego, Calif. : 1997), 2007
The inductance of a radiofrequency coil determines its compatibility with a given NMR probe circu... more The inductance of a radiofrequency coil determines its compatibility with a given NMR probe circuit. However, calculation (or estimation) of inductance for radiofrequency coils of dimensions suitable for use in an NMR probe is not trivial, particularly for flat-coils. A comparison of a number of formulae for calculation of inductance is presented through the use of a straightforward inductance measurement circuit. This technique relies upon instrumentation available in many NMR laboratories rather than upon more expensive and specialized instrumentation often utilized in the literature. Inductance estimation methods are suggested and validated for both flat-coils and solenoids. These have proven very useful for fabrication of a number of new coils in our laboratory for use in static solid-state NMR probes operating at (1)H frequencies of 300 and 600MHz. Solenoidal coils with very similar measured and estimated inductances having inner diameters from 1 to 5mm are directly compared as...
The reovirus p10 fusion-associated small transmembrane (FAST) proteins are the smallest known mem... more The reovirus p10 fusion-associated small transmembrane (FAST) proteins are the smallest known membrane fusion proteins, and evolved specifically to mediate cell-cell, rather than virus-cell, membrane fusion. The 36-40-residue ectodomains of avian reovirus (ARV) and Nelson Bay reovirus (NBV) p10 contain an essential intramolecular disulfide bond required for both cell-cell fusion and lipid mixing between liposomes. To more clearly define the functional, biochemical and biophysical features of this novel fusion peptide, synthetic peptides representing the p10 ectodomains of ARV and NBV were analyzed by solution-state NMR spectroscopy, circular dichroism spectroscopy, fluorescence spectroscopy-based hydrophobicity analysis, and liposome binding and fusion assays. Results indicate that disulfide bond formation promotes exposure of hydrophobic residues, as indicated by bis-ANS binding and time-dependent peptide aggregation under aqueous conditions, implying the disulfide bond creates a s...
A holder allowing rotation of a flat coil relative to the static magnetic field (B0) of a nuclear... more A holder allowing rotation of a flat coil relative to the static magnetic field (B0) of a nuclear magnetic resonance (NMR) spectrometer over the range of orientations from parallel to perpendicular has been built and incorporated into a two-channel wide line static solid-state NMR probe. Radiofrequency induction by the coil is perpendicular to B0 for all rotational angles. Such a
Spider silk is one of nature&... more Spider silk is one of nature's most remarkable biomaterials due to extraordinary strength and toughness not found in today's synthetic materials. Of the seven types of silk, wrapping silk (AcSp1) is the most extensible of the types of silks and has no sequence similarity to the other types. Here we report the chemical shifts for the AcSp1 199 amino acid protein repeat unit and its anticipated secondary structure based on secondary chemical shifts.
+ /H + exchanger) is a ubiquitously ex- pressed integral membrane protein that regulates intracel... more + /H + exchanger) is a ubiquitously ex- pressed integral membrane protein that regulates intracellular pH by removing a proton in exchange for an extracellular sodium ion. Of the nine known isoforms of the mammalian NHEs, the firstisoformdiscovered(NHE1)isthemostthoroughlycharacter- ized. NHE1 is involved in numerous physiological processes in mammals, including regulation of intracellular pH, cell-volume control, cytoskeletal organization, heart disease and cancer. NHE comprises two domains: an N-terminal membrane domain that functions to transport ions, and a C-terminal cytoplasmic regul- atory domain that regulates the activity and mediates cytoskeletal interactions.AlthoughtheexactmechanismoftransportbyNHE1 remainselusive,recentstudieshaveidentifiedaminoacidresidues that are important for NHE function. In addition, progress has been made regarding the elucidation of the structure of NHEs. Specifically, the structure of a single TM (transmembrane) seg- ment from NHE1 has been sol...
Rhomboid proteases are integral membrane serine proteases that catalyze peptide bond hydrolysis i... more Rhomboid proteases are integral membrane serine proteases that catalyze peptide bond hydrolysis in biological membranes. Little is currently known about the interaction of enzyme and substrate. Coarse-grained molecular dynamics simulations in hydrated lipid bilayers are employed herein to study the interaction of the E. coli rhomboid protease GlpG (ecGlpG) with the transmembrane domain (TMD) of the substrate Spitz. Spitz does not associate with ecGlpG exclusively at the putative substrate gate near TMD 5. Instead, there are six prominent and stable interaction sites, including one between TMDs 1 and 3, with the closest enzyme-substrate proximity occurring at the ends of helical TMDs or in loops. Bilayer thinning is observed proximal to ecGlpG, but there is no evidence of additional thinning of the bilayer upon interaction with substrate. We suggest that the initial interaction between enzyme and substrate, or substrate capture event, is not limited to a single site on the enzyme, and may be driven by juxtamembrane electrostatic interactions. The findings are of additional interest because catalytically inactive rhomboids (iRhoms) are now known to interact with the substrates of their catalytically active counterparts and to antagonize the enzyme-driven pathways.
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Papers by Jan Rainey