Showing papers in "Biochemical Journal in 1963"

The preparation of 131i-labelled human growth hormone of high specific radioactivity

Abstract: A simple and rapid method is presented for the preparation of I/sup 131/- labeled human growth hormone of high specific radioactivity (240-300 mu C/ mu g). Low amounts of carrierfree I/sup 131/ iodide (2 mC) are allowed to react, without prior treatment, with small quantities of protein (5 mu g) in a highyield reaction (approx. 70% transfer of I/sup 131/ to protein). The degree of chemical substitution is minimized (0.5- 1.0 atom of iodine/molecule of protein) by the use of carrier-free I/sup 131/ iodide. The I/sup 131/-labeled hormone (up to 300 mu C/ mu g) contains no detectable degradation products and is immunologically identical with the unlabeled hormone. The loss of immunological reactivity at high specific radioactivities or at high levels of chemical substitution with STAI/sup 127/!iodine is demonstrated. (auth)

Further studies on the alkylation of nucleic acids and their constituent nucleotides

Abstract: Prothidium caused gross loss of motility and clumping of all trypanosomes. Comparison of the findings for trypanosomes with those for other cells previously studied shows that T. Iewi8i iS very similar electrophoretically to freely circulating cells such as lymphocytes and tumour cells. On the other hand, the surface properties of the blood-stream form of T. rhodesiense are different electrophoretically from any other freely circulating cell so far studied. If the surface of these cells does contain carboxyl groups, the previous hypothesis that all cells carrying surface carboxyl groups will be adhesive to phagocytes will need modification (Bangham & Pethica, 1960). The absence of net charge on the freelycirculating blood-stream form of T. rhodesiense is evidence that van der Waals forces play a very minor role in cell adhesion (Pethica, 1961), although the flagellate motility may adversely influence the adhesion process.

The arrangement of the peptide chains in γ-globulin

Abstract: Hardwick, D.C. & Linzell, J.L. (1960). J. Physiol. 154, 547. Hardwick, D. C., Linzell, J. L. & Price, S. M. (1961). Biochem. J. 80 37. Huggett, A. St G. & Nixon, D. A. (1957). Lancet, ii, 368. Kisin, I. E. & Tsaturov, Y. L. (1961). Bull. exp. Biol. Med., U.S.S.R. (Engl. trans.) 50, 864. Kleiber, M. (1954). Rev. Canad. Biol. 13, 333. Kleiber, M., Black, A. L., Brown, M. A., Baxter, C. F., Luick, J. R. & Stadtman, F. H. (1955). Biochim. biophys. Acta, 17, 252. Lindsay, D. B. (1959). Vet. Rev. 5, 103. Lintzel, W. (1934). Lait, 14, 1125. Linzell, J. L. (1960). J. Physiol. 153, 492. Lucas, J. M., Kaneko, J. J., Hirohara, K. & Kleiber, M. (1959). Agric. Fd Chem. 7, 638. Luick, J. R. (1960). J. Dairy Sci. 43, 1344. Luick, J. R. & Kleiber, M. (1961). Amer. J. Physiol. 200, 1327. McArdle, B. A. (1955). Biochem. J. 60, 647. Macy, I. G., Kelly, H. J. & Sloan, R. E. (1953). The Composition of Milks. Washington: National Academy of Science National Research Council, Publication no. 254. Nelson, N. (1944). J. biol. Chem. 153, 375. Peirce, E. C. (1960). J. thorac. Surg. 39, 438. Popjak, G., Folley, S. J. & French, T. H. (1951). Biochem. J. 48, 44. Popjak, G., French, T. H., Hunter, G. D. & Martin, A. J. P. (1951). Biochem. J. 48, 612. Riis, P. M., Luick, J. R. & Kleiber, M. (1960). Amer. J. Physiol. 198, 45. Somogyi, M. (1952). J. biol. Chem. 195, 19. Spencer, A. F. & Lowenstein, J. M. (1962). J. biol. Chem. 237, 3640. Srere, P. A. & Lipmann, F. (1953). J. Amer. chem. Soc. 75, 4874. Tombropoulos, E. G. & Kleiber, M. (1961). Biochem. J. 80, 414. Utter, M. F. (1959). Ann. N. Y. Acad. Sci. 72, 451. Utter, M. F. & Keech, D. B. (1960). J. biol. Chem. 235, Pc17. Weinman, E. O., Strisower, E. H. & Chaikoff, I. L. (1957). Physiol. Rev. 37, 252. Wood, H. G., Gillespie, R., Hansen, R. G., Wood, W. A. & Hardenbrooke, H. J. (1959). Biochem. J. 73, 694.

The frequency of errors in protein biosynthesis

Abstract: Anson, M. L. (1938). J. gen. Physiol. 22, 79. Brown, P. C., Consden, R. & Glynn, L. E. (1958). Ann. Rheum. Di8. 17, 196. Dawson, A. B. (1946). Amer. J. Anat. 79, 241. Dingle, J. T. (1962). Proc. roy. Soc. Med. 55, 109. Frankland, D. M. & Wynn, C. H. (1962). Biochem. J. 85, 276. Harkness, M. L. R. & Harkness, R. D. (1954). J. Physiol. 123, 492. Harkness, R. D. & Moralee, B. E. (1956). J. Physiol. 132, 502. Lohel, B. L. & Deane, H. WV. (1962). Endocrinology, 70, 567. Lowry, 0. H., Gilligan, D. R. & Katersky, E. M. (1941). J. biol. Chem. 139, 795. Maher, J. A. (1959). Arch. Path. 67, 175. Mavromati, L. (1950). Ann. Endocr., Paris, 11, 148. Montfort, I. & Perez-Tamayo, R. (1961). Lab. Invest. 10, 1240. Morrione, T. G. & Seifter, S. (1962). J. exp. Med. 115, 357. Neuman, R. E. & Logan, M. A. (1950). J. biol. Chem. 186, 549. Partridge, S. M. & Davis, H. F. (1955). Biochem. J. 61, 21. Schwalm, H. & Cretius, K. (1958). Arch. Gyndk. 191, 271. Troll, W. & Lindsley, J. (1955). J. biol. Chem. 215, 655. Weiss, L. (1962). Biochem. J. 83, 18P. Woessner, J. F., jun. (1961). Arch. Biochem. Biophys. 93, 440. Woessner, J. F., jun. (1962a). Biochem. J. 83, 304. Woessner, J. F., jun. (1962b). J. Geront. 17, 453 (abstr.). Woessner, J. F., jun. & Brewer, T. H. (1960). Fed. Proc. 19, 335.

Studies on the mode of action of excess of vitamin A. 6. Lysosomal protease and the degradation of cartilage matrix.

Abstract: complex ofthematrix, andthatliberation ofother lysosomal hydrolytic enzymesmightcausethecellular changes observed incartilage cultivated inthepresence of thevitamin; thishypothesis was supported by observations onthespecificity oftheaction ofthe vitamin, bothinorganculture andontheisolated lysosomes (Fell, Dingle& Webb,1962).Recent experiments ontheeffect ofthevitamin onthecell membraneoferythrocytes (Dingle & Lucy,1962), andonmitochondria (J.A.Lucy,M.Luscombe& J.T.Dingle, unpublished work),provide further evidence thatonesite ofvitamin A action isatthe lipoprotein membranes ofcells andtheir organelles. Thoughtheevidence sofarobtained favoured thishypothesis concerning themodeofaction of vitamin A oncartilage, threeimportant pieces of information werestill lacking. (1)Although normal cartilaginous rudiments hadbeenfoundtocontain a protease, withanacidpHoptimum, thatwhenfreed by treatmentwithhypo-osmotic solutions can degrade thematrix(Lucyetal.1961), andthough therewas increased lysis oftheplasmaclotby cartilaginous rudiments growninthepresence of excess ofvitamin A (Dingle, Lucy& Fell, 1961), the liberation ofanacidprotease bythevitamin Atreated explants had notbeenclearly demonstrated. (2)Itwasnotknownwhether thevitamin could release theenzymesfromintracellular particles isolated fromcartilage, thoughithadbeenshownto liberate aprotease fromlysosomes isolated fromrat liver (Dingle, 1961). (3)Itwasnotknownwhether acidhydrolases fromisolated intracellular particles coulddegrade cartilage rudiments. Thesethree points havenowbeeninvestigated.

The interaction between polysaccharides and other macromolecules. 4. The osmotic pressure of mixtures of serum albumin and hyaluronic acid

Abstract: Ogston & Phelps (1961) found that hyaluronic acid markedly affects the partition of diffusible macromolecules between solutions of the polysaccharide and buffer. The authors explain the phenomenon as a steric exclusion of macromolecular solutes from solutions containing randomly coiled hyaluronic acid chains. From these results one can expect that hyaluronic acid and similar substances would exert a significant influence on the thermodynamic properties of solutions containing other macromolecules. The investigation reported in the present paper shows that solutions containing both hyaluronic acid and serum albumin have osmotic pressures in excess of the sum of osmotic pressures of solutions containing hyaluronic acid and serum albumin separately at the same concentrations. The influence of the 'effective volume' of a solute of finite concentration on the osmotic pressure of a solution has been treated by a number of authors (see, for example, Scatchard, 1946; Doty & Edsall, 1951; Edsall, 1953; Flory, 1953). Christiansen (1960), using the experimental results of Jensen & Marcker (1958), discussed the phenomenon in hyaluronic acid solutions. All these treatments, however, have been limited to the problem of binary systems containing the solvent and a single species of macromolecules. Ogston (1962) discussed from a theoretical point of view the thermodynamic properties of ternary systems with special reference to buffer solutions containing hyaluronic acid and serum albumin. The study of this problem was approached independently by the present authors. Experimental work was planned and carried out by one of us (T. C. L.) and a theoretical treatment of the thermodynamic properties of systems containing hyaluronic acid and serum albumin was made by the other (Ogston, 1962). The present paper combines the two approaches.

Metabolism of d-glucuronolactone in mammalian systems. Identification of d-glucaric acid as a normal constituent of urine.

Abstract: The inhibition of P-glucuronidase by mamResearch Unit, Aberdeen Royal Infirmary, to whom the malian urine has been noted by a number of author is indebted. Fresh samples of cat urine were workers. This complicates assay of the enzyme in supplied by Dr A. J. Carr, Pathology Department, Aberurine, and the use of added *-e to deen University; 24 hr. urines of rat and guinea pig, and urine, and the us of -glucuronidase 1 samples of sheep, pig and calf urines, were collected from lhberate excretedl phenols and alcohols' includm' let xo s, i ng animals maintained on normal diets in metabolism cages. steroids, from the fl-D-glucosiduronic acid conThe rat and guinea-pig exereta were separated by filtration, jugates. P-Glucuronidase assay is normally conand the faeces washed with water, before the combined ducted with a chromogenic substrate, the enzymic urine and washings were tested. Urines were stored at 0' if hydrolysis of which may be diminished in the examined within 24 hr. of collection, or otherwise at 20°. presence of urine which contains competing subTreatment of urine. The following standard procedure strates of different affinities (see Levvy & Marsh, was used in testing the inhibitory powers of mammalian 1954). In addition to natural substrates for p urine. Crude urine (pH 4-5-7-0) was treated (a) at 1000 glucuronidase, however, mammnalian urine confor 40 min. after adjustment with 3N-HCl to pH 2-0-2-2, tains trueenzymeinhibitrs Abul-Fadl(1957) followed by readjustment with NaOH to pH 4-0-45, or tamns true enzyme inhibitors. Abul-Fadl (1957) (b) at 100° for 15 min. after adjustment with 0-1 N-NaOH to found both dialysable and non-dialysable material pH 7-5-8-0, follQwed by readjustment with HCI to pH 60in human urine which was inhibitory to /-gluc6-5. The inhibitory power was maximal after treatment uronidase; the dialysable inhibitor was stable to (a) and minimal after treatment (b). Either treatment was treatment with hot acid. Similar inhibitory fracsufficient to rid the urine of ,B-glucuronidase activity, and tions were present in rabbit urine (Conzelman & after the readjustments there was no interference with the Crout, 1961). pH of the subsequent enzyme assay. Heating at a pH Mammalian /3-glucuronidase is inhibited by above 2-5 was inadequate for development of maximum heavy-metal ions (Levvy & Marsh, 1957a, b; inhibitory power of acidified urines, for there was then a

A polymer containing glucose and aminohexuronic acid isolated from the cell walls of micrococcus lysodeikticus

Abstract: The surveys of Cummins & Harris (1956, 1958) established that the cell walls of Gram-positive bacteria all contain the amino sugars glucosamine and muramic acid (3-O-a.-carboxyethylglucosamine) and the amino acids alanine, glutamic acid and either lysine or aci-diaminopimelic acid. In some species either glycine or aspartic acid was also present, whereas the other amino acids found in proteins were usually absent. The component of cell-wall structure containing these compounds has been found in all bacteria so far examined and has been given the name mucopeptide (Perkins & Rogers, 1959). In addition, various non-nitrogenous sugars have been found in the cell walls of some strains, but the place of these components in the structure of the wall is not always understood. Often the complex polyol phosphate polymers known as teichoic acids (Baddiley, 1959) are present in the cell walls, and these compounds may include sugar residues glycosidically linked to the polyol. For instance, glucose occurs in the ribitol teichoic acid isolated from the cell walls of Bacillus subtilis (Armstrong, Baddiley & Buchanan, 1960). Sometimes, however, sugars are found that are not part of the teichoic acid but which are present in polymers independent of both teichoic acid and mucopeptide. Examples are the rhamnose-glucosamine polymer found in Streptococcus faecalis (Ikawa, 1961) and group A haemolytic streptococci (McCarty, 1960), and the teichuronic acid, a polymer of N-acetylgalactosamine and glucuronic acid, found in the cell walls of B. subtilis (Janczura, Perkins & Rogers, 1961). From previous work it was known that the cell walls of Micrococcus lysodeiktic contain no phosphorus (and hence no teichoic acid) and consist mainly of glucosamine, muramic acid, alanine, glutamic acid, lysine, glycine and glucose. A critical analytical study has shown that these constituents together account for 93 % of the dry weight of the walls, after allowance has been made for the uptake of water during hydrolysis (Czerkawski, Perkins & Rogers, 1963). Nevertheless, certain discrepancies were brought to light, parti-