Regulation of syntaxin1A–munc18 complex for SNARE pairing in HEK293 cells

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65811/1/jphysiol.2004.067249.pd

Ultra-cold methods for polarized atomic hydrogen

Using the ultra-cold electron-spin-polarized atomic hydrogen technique, one can produce a slow monochromatic beam for use as a polarized jet target. We will first review the development of the ultra-cold technique and then discuss the recent progress on Michigan’s Mark-II ultra-cold proton-spin-polarized hydrogen jet target. © 1998 American Institute of Physics.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87438/2/119_1.pd

Michigan ultra-cold polarized atomic hydrogen jet target

To study spin effects in high energy collisions, we are developing an ultra-cold high-density jet target of proton-spin-polarized hydrogen atoms. The target uses a 12 Tesla magnetic field and a 0.3 K separation cell coated with superfluid helium-4 to produce a slow monochromatic electron-spin-polarized atomic hydrogen beam, which is then focused by a superconducting sextupole into the interaction region. In recent tests, we studied a polarized beam of hydrogen atoms focused by the superconducting sextupole into a compression tube detector, which measured the polarized atoms’ intensity. The Jet produced, at the detector, a spin-polarized atomic hydrogen beam with a measured intensity of about 2.8⋅1015 H s−12.8⋅1015Hs−1 and a FWHM area of less than 0.13 cm2. This intensity corresponds to a free jet density of about 1⋅1012 H cm−31⋅1012Hcm−3 with a proton polarization of about 50%. When the transition RF unit is installed, we expect a proton polarization higher than 90%. © 2001 American Institute of Physics.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87742/2/856_1.pd

Polarized atomic hydrogen beam studies in the Michigan ultra-cold jet

Studies of an ultra-cold jet of polarized hydrogen atoms are described. This atomic beam is formed by the acceleration of cold (0.3 K) atoms emerging from a region of high magnetic field (12 T). The maximum measured density was about 1012 atoms cm−3.1012atomscm−3. The beam’s full width half maximum size was less than 4 mm. © 2000 American Institute of Physics.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87560/2/674_1.pd

Positional Cloning of a Type 2 Diabetes Quantitative Trait Locus; Tomosyn-2, a Negative Regulator of Insulin Secretion

We previously mapped a type 2 diabetes (T2D) locus on chromosome 16 (Chr 16) in an F2 intercross from the BTBR T (+) tf (BTBR) Lepob/ob and C57BL/6 (B6) Lepob/ob mouse strains. Introgression of BTBR Chr 16 into B6 mice resulted in a consomic mouse with reduced fasting plasma insulin and elevated glucose levels. We derived a panel of sub-congenic mice and narrowed the diabetes susceptibility locus to a 1.6 Mb region. Introgression of this 1.6 Mb fragment of the BTBR Chr 16 into lean B6 mice (B6.16BT36–38) replicated the phenotypes of the consomic mice. Pancreatic islets from the B6.16BT36–38 mice were defective in the second phase of the insulin secretion, suggesting that the 1.6 Mb region encodes a regulator of insulin secretion. Within this region, syntaxin-binding protein 5-like (Stxbp5l) or tomosyn-2 was the only gene with an expression difference and a non-synonymous coding single nucleotide polymorphism (SNP) between the B6 and BTBR alleles. Overexpression of the b-tomosyn-2 isoform in the pancreatic β-cell line, INS1 (832/13), resulted in an inhibition of insulin secretion in response to 3 mM 8-bromo cAMP at 7 mM glucose. In vitro binding experiments showed that tomosyn-2 binds recombinant syntaxin-1A and syntaxin-4, key proteins that are involved in insulin secretion via formation of the SNARE complex. The B6 form of tomosyn-2 is more susceptible to proteasomal degradation than the BTBR form, establishing a functional role for the coding SNP in tomosyn-2. We conclude that tomosyn-2 is the major gene responsible for the T2D Chr 16 quantitative trait locus (QTL) we mapped in our mouse cross. Our findings suggest that tomosyn-2 is a key negative regulator of insulin secretion

Analyzing Power AnA_n in High P-Transverse Squared Proton-Proton Elestic Scattering

This is a proposal to measure the Analyzing Power $A_n$ in Proton-Proton Elestic Scattering at High P-Transverse Squared of 1 to 12 (GeV/c)² using a 120 GeV unpolarized extracted proton beam from Fermilab's Main Injector starting in 2001

SNARE-catalyzed Fusion Events Are Regulated by Syntaxin1A–Lipid Interactions

Membrane fusion is a process that intimately involves both proteins and lipids. Although the SNARE proteins, which ultimately overcome the energy barrier for fusion, have been extensively studied, regulation of the energy barrier itself, determined by specific membrane lipids, has been largely overlooked. Our findings reveal a novel function for SNARE proteins in reducing the energy barrier for fusion, by directly binding and sequestering fusogenic lipids to sites of fusion. We demonstrate a specific interaction between Syntaxin1A and the fusogenic lipid phosphatidic acid, in addition to multiple polyphosphoinositide lipids, and define a polybasic juxtamembrane region within Syntaxin1A as its lipid-binding domain. In PC-12 cells, Syntaxin1A mutations that progressively reduced lipid binding resulted in a progressive reduction in evoked secretion. Moreover, amperometric analysis of fusion events driven by a lipid-binding–deficient Syntaxin1A mutant (5RK/A) demonstrated alterations in fusion pore dynamics, suggestive of an energetic defect in secretion. Overexpression of the phosphatidic acid–generating enzyme, phospholipase D1, completely rescued the secretory defect seen with the 5RK/A mutant. Moreover, knockdown of phospholipase D1 activity drastically reduced control secretion, while leaving 5RK/A-mediated secretion relatively unaffected. Altogether, these data suggest that Syntaxin1A–lipid interactions are a critical determinant of the energetics of SNARE-catalyzed fusion events