Identification of N-terminal regions of wheat leaf ferredoxin NADP+ oxidoreductase important for interactions with ferredoxin.

Wheat leaves contain two isoproteins of the photosynthetic ferredoxin:NADP(+) reductase (pFNRI and pFNRII). Truncated forms of both enzymes have been detected in vivo, but only pFNRII displays N-terminal length-dependent changes in activity. To investigate the impact of N-terminal truncation on interaction with ferredoxin (Fd), recombinant pFNRII proteins, differing by deletions of up to 25 amino acids, were generated. During purification of the isoproteins found in vivo, the longer forms of pFNRII bound more strongly to a Fd affinity column than did the shorter forms, pFNRII(ISKK) and pFNRII[N-2](KKQD). Further truncation of the N-termini resulted in a pFNRII protein which failed to bind to a Fd column. Similar k(cat) values (104-140 s(-1)) for cytochrome c reduction were measured for all but the most truncated pFNRII[N-5](DEGV), which had a k(cat) of 38 s(-1). Stopped-flow kinetic studies, examining the impact of truncation on electron flow between mutant pFNRII proteins and Fd, showed there was a variation in k(obs) from 76 to 265 s(-1) dependent on the pFNRII partner. To analyze the sites which contribute to Fd binding at the pFNRII N-terminal, three mutants were generated, in which a single or double lysine residue was changed to glutamine within the in vivo N-terminal truncation region. The mutations affected binding of pFNRII to the Fd column. Based on activity measurements, the double lysine residue change resulted in a pFNRII enzyme with decreased Fd affinity. The results highlight the importance of this flexible N-terminal region of the pFNRII protein in binding the Fd partner.

Measurement of the nonlinear coefficient of orientation-patterned GaAs and demonstration of highly efficient second-harmonic generation.

Quasi-phase-matched (QPM) GaAs structures, 0.5 mm thick, 10 mm long, and with 61-mum grating periods, were grown by a combination of molecular-beam epitaxy and hydride vapor phase epitaxy. These were characterized by use of mid-IR second-harmonic generation (SHG) with a ZnGeP(2) (ZGP) optical parametric oscillator as a pump source. The SHG efficiencies of QPM GaAs and QPM LiNbO(3) were directly compared, and a ratio of nonlinear coefficients d(14)(GaAs)/d(33) (LiNbO(3))=5.01+/-0.3 was found at 4.1-mum fundamental wavelength. For input pulse energies as low as 50muJ and approximately 60-ns pulse duration, an internal SHG conversion efficiency of 33% was measured in QPM GaAs.

Difference frequency generation of 8-microm radiation in orientation- patterned GaAs.

First-order quasi-phase-matched difference frequency generation of narrowband tunable mid-infrared light is demonstrated in orientation-patterned GaAs. The all-epitaxial orientation-patterned crystal is fabricated by a combination of molecular beam epitaxy and hydride vapor phase epitaxy. Lasers at 1.3 and 1.55 microm were mixed to give an idler output at 8 microm, with power and wavelength tuning consistent with theoretical estimates, indicating excellent material uniformity over the 19-mm-long and 500-microm-thick device.

Presence of phosphorylcholine on a filarial nematode protein influences immunoglobulin G subclass response to the molecule by an interleukin-10-dependent mechanism.

The filarial nematode product ES-62 contains phosphorylcholine (PC) covalently attached to N-linked glycans. ES-62 induced high levels of immunoglobulin G1 (IgG1) antibodies, but no IgG2a, to non-PC epitopes of the molecule following subcutaneous injection into BALB/c mice. Conversely, mice given ES-62 lacking PC demonstrated significant production of both IgG subclasses. Thus, PC appears to block production of IgG2a antibodies to other epitopes on the parasite molecule. A role for interleukin-10 (IL-10) in this effect was shown by the ability of IL-10(-/-) mice to make an IgG2a antibody response to non-PC epitopes of ES-62.