Location of alkali metal binding sites in endothelin A selective receptor antagonists, cyclo(D-Trp-D-Asp-Pro-D-Val-Leu) and cyclo(D-Trp-D-Asp-Pro-D-Ile-Leu), from multistep collisionally activated decompositions

We previously showed by using mass spectrometry that endothelin A selective receptor antagonists BQ123 and JKC301 form novel coordination compounds with sodium ions. This property may underlie the ability of an ET(A) antagonist to induce net tubular sodium reabsorption in the proximal tubule cells and reverse acute renal failure induced by severe ischemia. We have now defined the metal binding sites on BQ123 and JKC301 by subjecting the metal-containing peptides to multiple stages of collisionally activated decomposition (CAD) in an ion trap mass spectrometer. When submitted to low-energy CAD, the ring opens at the Asp-Pro amide bond. The metal ion, which bonds, inter alia, to the carbonyl oxygen of the proline residue, acts as a fixed charge site, and directs a charge-remote, sequence-specific fragmentation of the ring-opened peptide. Amino acid residues are sequentially cleaved from the C-terminal end, and the terminal aziridinone structure moves one step toward the N-terminus with each C-terminal amino acid residue removed. These observations are the basis of a new method to sequence cyclic peptides. Amino acid residues are observed as sets of three ions, a*(n)PD, b*(n)PD and c*(n)PD where n is the number of amino acid residues in the peptide.     

Identification of a set of genes showing regionally enriched expression in the mouse brain

Background

We have recorded responses from single neurons in murine visual cortex to determine the effectiveness of the input from the two murine cone photoreceptor mechanisms and whether there is any unique selectivity for cone inputs at this higher region of the visual system that would support the possibility of colour vision in mice. Each eye was stimulated by diffuse light, either 370 (strong stimulus for the ultra-violet (UV) cone opsin) or 505 nm (exclusively stimulating the middle wavelength sensitive (M) cone opsin), obtained from light emitting diodes (LEDs) in the presence of a strong adapting light that suppressed the responses of rods.

Results

Single cells responded to these diffuse stimuli in all areas of striate cortex. Two types of responsive cells were encountered. One type (135/323 – 42%) had little to no spontaneous activity and responded at either the on and/or the off phase of the light stimulus with a few impulses often of relatively large amplitude. A second type (166/323 – 51%) had spontaneous activity and responded tonically to light stimuli with impulses often of small amplitude. Most of the cells responded similarly to both spectral stimuli. A few (18/323 – 6%) responded strongly or exclusively to one or the other spectral stimulus and rarely in a spectrally opponent manner.

Conclusion

Most cells in murine striate cortex receive excitatory inputs from both UV- and M-cones. A small fraction shows either strong selectivity for one or the other cone mechanism and occasionally cone opponent responses. Cells that could underlie chromatic contrast detection are present but extremely rare in murine striate cortex.     

Multistep tandem mass spectrometry for Sequencing Cyclic Peptides in an Ion-Trap Mass Spectrometer

Collisionally activated decomposition (CAD) of a protonated cyclic peptide produces a superposition spectrum consisting of fragments produced following random ring opening of the cyclic peptide to give a set of acylium ions (or isomeric equivalents) of the same m/z. Assignment of the correct sequence is often difficult owing to lack of selectivity in the ring opening. A method is presented that utilizes multiple stages of CAD experiments in an electrospray ion-trap mass spectrometer to sequence cyclic peptides. A primary acylium ion is selected from the primary product-ion spectrum and subjected to several stages of CAD. Amino-acid residues are sequentially removed, one at each stage of the CAD, from the C-terminus, until a b2 ion is reached. Results are presented for seven cyclic peptides, ranging in sizes from four to eight amino-acid residues. This method of sequencing cyclic peptides eliminates ambiguities encountered with other MS/MS approaches. The power of the strategy lies in the capability to execute several stages of CAD upon a precursor ion and its decomposition products, allowing the cyclic peptide to be sequenced in an unambiguous, stepwise manner.     

Effect of substrate temperature on electrical and magnetic properties of epitaxial La1−x Pb x MnO3 films

Epitaxial La_1−x Pb _x MnO₃ (LPMO) thin films, grown on (100) SrTiO₃ substrates by laser ablation technique at different temperatures between 600 and 850°C, have been characterized for electrical and magnetic properties. The temperature dependence of resistivity showed that the metal-insulator transition temperature (T _MI) decreases with increasing substrate temperature, which has been attributed to decrease in Pb content in the filsm. The YBa₂Cu₃O _x /La_1−x MnO₃ heterostructures, exhibiting both superconductivity and ferromagnetism, have been fabricated.     

A Criterion for Finite Topological Determinacy of Map-Germs

Let f, g: (Rⁿ, 0) (R^p, 0) be two C map-germs. Then f and gare C⁰-equivalent if there exist homeomorphism-germs h and lof (Rⁿ, 0) and (R^p, 0) respectively such that g = l f h^–1.Let k be a positive integer. A germ f is k-C⁰-determined ifevery germ g with j^k g(0) = j^k f(0) is C⁰-equivalent to f. Moreover,we say that f is finitely topologically determined if f is k-C⁰-determinedfor some finite k. We prove a theorem giving a sufficient conditionfor a germ to be finitely topologically determined. We explainthis condition below. Let N and P be two C manifolds. Consider the jet bundle J^k(N,P) with fiber J^k(n, p). Let z in J^k(n, p) and let f be suchthat z = j^kf(0). Define Whether (f) < k depends only on z, not on f. We can thereforedefine the set Let W^k(N, P) be the subbundle of J^k(N, P) with fiber W^k(n, p).Mather has constructed a finite Whitney (b)-regular stratificationS^k(n, p) of J^k(n, p) – W^k(n, p) such that all strata aresemialgebraic and K-invariant, having the property that if S^k(N,P) denotes the corresponding stratification of J^k(N, P) –W^k(N, P) and f C(N, P) is a C map such that j^kf is multitransverseto S^k(N, P), j^kf(N) W^k(N, P) = and N is compact (or f is proper),then f is topologically stable. For a map-germ f: (Rⁿ, 0) (R^p, 0), we define a certain ojasiewiczinequality. The inequality implies that there exists a representativef: U R^p such that j^kf(U – 0) W^k (Rⁿ, R^p = and suchthat j^kf is multitransverse to S^k (Rⁿ, R^p) at any finite setof points S U – 0. Moreover, the inequality controlsthe rate j^kf becomes non-transverse as we approach 0. We showthat if f satisfies this inequality, then f is finitely topologicallydetermined. 1991 Mathematics Subject Classification: 58C27.