Generation of cDNA probes directed by amino acid sequence: cloning of urate oxidase

Urate oxidase (E.C. 1.7.3.3) catalyzes the oxidation of uric acid to allantoin in most mammals except humans and certain primates. The amino-terminal amino acid sequence for porcine urate oxidase was determined and used in a novel procedure for generating complementary DNA (cDNA) probes to this amino acid sequence. The procedure is based on the polymerase chain reaction and utilizes mixed oligonucleotide primers complementary to the reverse translation products of an amino acid sequence. This rapid and simple cDNA cloning procedure is generally applicable and requires only a partial amino acid sequence. A cDNA probe developed by this procedure was used to isolate a full-length porcine urate oxidase cDNA and to demonstrate the presence of homologous genomic sequences in humans.     

Composition and structure of the martian atmosphere: preliminary results from viking 1

Results from the aeroshell-mounted neutral mass spectrometer on Viking I indicate that the upper atmosphere of Mars is composed mainly of CO(2) with trace quantities of N(2), Ar, O, O(2), and CO. The mixing ratios by volume relative to CO(2) for N(2), Ar, and O(2) are about 0.06, 0.015, and 0.003, respectively, at an altitude near 135 kilometers. Molecular oxygen (O(2)(+)) is a major component of the ionosphere according to results from the retarding potential analyzer. The atmosphere between 140 and 200 kilometers has an average temperature of about 180 degrees +/- 20 degrees K. Atmospheric pressure at the landing site for Viking 1 was 7.3 millibars at an air temperature of 241 degrees K. The descent data are consistent with the view that CO(2) should be the major constituent of the lower martian atmosphere.     

Replacements of Pro86 in phage T4 lysozyme extend an alpha-helix but do not alter protein stability

To investigate the relation between protein stability and the predicted stabilities of individual secondary structural elements, residue Pro86 in an alpha-helix in phage T4 lysozyme was replaced by ten different amino acids. The x-ray crystal structures of seven of the mutant lysozymes were determined at high resolution. In each case, replacement of the proline resulted in the formation of an extended alpha-helix. This involves a large conformational change in residues 81 to 83 and smaller shifts that extend 20 angstroms across the protein surface. Unexpectedly, all ten amino acid substitutions marginally reduce protein thermostability. This insensitivity of stability to the amino acid at position 86 is not simply explained by statistical and thermodynamic criteria for helical propensity. The observed conformational changes illustrate a general mechanism by which proteins can tolerate mutations.     

Triton's Geyser-Like Plumes: Discovery and Basic Characterization

At least four active geyser-like eruptions were discovered in Voyager 2 images of Triton, Neptune's large satellite. The two best documented eruptions occur as columns of dark material rising to an altitude of about 8 kilometers where dark clouds of material are left suspended to drift downwind over 100 kilometers. The radii of the rising columns appear to be in the range of several tens of meters to a kilometer. One model for the mechanism to drive the plumes involves heating of nitrogen ice in a subsurface greenhouse environment; nitrogen gas pressurized by the solar heating explosively vents to the surface carrying clouds of ice and dark partides into the atmosphere. A temperature increase of less than 4 kelvins above the ambient surface value of 38 +/- 3 kelvins is more than adequate to drive the plumes to an 8-kilometer altitude. The mass flux in the trailing clouds is estimated to consist of up to 10 kilograms of fine dark particles per second or twice as much nitrogen ice and perhaps several hundred or more kilograms of nitrogen gas per second. Each eruption may last a year or more, during which on the order of a tenth of a cubic kilometer of ice is sublimed.