Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants

The crystallization of three catalytically inactive mutants of penicillin Vacylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide threedimensional structures of precursor PVA forms, plus open a route to the study of enzyme-substrate complexes for this industrially important enzyme

Multi-transition study and new detections of class II methanol masers

We have used the ATNF Mopra antenna and the SEST antenna to search in the directions of several class II methanol maser sources for emission from six methanol transitions in the frequency range 85-115 GHz. The transitions were selected from excitation studies as potential maser candidates. Methanol emission at one or more frequencies was detected from five of the maser sources, as well as from Orion KL. Although the lines are weak, we find evidence of maser origin for three new lines in G345.01+1.79, and possibly one new line in G9.62+0.20. The observations, together with published maser observations at other frequencies, are compared with methanol maser modelling for G345.01+1.79 and NGC6334F. We find that the majority of observations in both sources are consistent with a warm dust (175 K) pumping model at hydrogen density ~10^6 cm^-3 and methanol column density ~5 x 10^17 cm^-2. The substantial differences between the maser spectra in the two sources can be attributed to the geometry of the maser region.Comment: 13 pages, 6 figures, Accepted for publication in MNRA

Correlation between X-ray Lightcurve Shape and Radio Arrival Time in the Vela Pulsar

We report the results of simultaneous observations of the Vela pulsar in X-rays and radio from the RXTE satellite and the Mount Pleasant Radio Observatory in Tasmania. We sought correlations between the Vela's X-ray emission and radio arrival times on a pulse by pulse basis. At a confidence level of 99.8% we have found significantly higher flux density in Vela's main X-ray peak during radio pulses that arrived early. This excess flux shifts to the 'trough' following the 2nd X-ray peak during radio pulses that arrive later. Our results suggest that the mechanism producing the radio pulses is intimately connected to the mechanism producing X-rays. Current models using resonant absorption of radio emission in the outer magnetosphere as a cause of the X-ray emission are explored as a possible explanation for the correlation.Comment: 6 pages, 5 figures, accepted by Ap

Immune status of recipients following bone marrow - Augmented solid organ transplantation

It has been postulated that the resident “passenger” leukocytes of hematolymphoid origin that migrate from whole organ grafts and subsequently establish systemic chimerism are essential for graft acceptance and the induction of donor-specific nonreactivity. This phenomenon was augmented by infusing 3 × 108 unmodified donor bone-marrow cells into 40 patients at the time of organ transplantation. Fifteen of the first 18 analyzable patients had sequential immunological evaluation over postoperative intervals of 5 to 17 months, (which included 7 kidney (two with islets), 7 liver (one with islets), and one heart recipient). The evolution of changes was compared with that in 16 kidney and liver nonmarrow controls followed for 4 to5 months. The generic immune reactivity of peripheral blood mononuclear cells (PBMC) was determined by their proliferative responses to mitogens (PHA, ConA). Alloreactivity was measured by the recipient mixed lymphocyte reaction (MLR) to donor and HLA-mis-matched third-party panel cells. Based on all 3 tests,the recipients were classified as donor-specific hyporeactive, intermediate, and responsive; patients who were globally suppressed made up a fourth category. Eight (53%) of the 15 marrow-treated recipients exhibited progressive modulation of donor-specific reactivity (3 hyporeactive and 5 intermediate) while 7 remained antidonor-responsive. In the nonmarrow controls, 2 (12.5%) of the 16 patients showed donor-specific hyporeactivity, 10 (62.5%) were reactive, and 4 (25%) studied during a CMV infection had global suppression of responsiveness to all stimuli. © 1995 by Williams and Wilkins