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Biomedical subjects

L Visser

Publications and source records attributed to L Visser.

108 records · Page 6Linked to original sources
PubMed1977

Histidine and lysine residues and the activity of phospholipase A2 from the venom of Bitis gabonica.

Chemical modification of phospholipase A2 (phosphatide 2-acyl-hydrolase, EC 3.1.1.4) from the venom of gaboon adder (Bitis gabonica) showed that histidine and lysine residues are essential for enzyme activity. Treatment with p-bromophenacyl bromide or pyridoxal 5'-phosphate resulted in the specific covalent modification of one histidine or a total of one lysine residue per molecule of enzyme, respectively, with a concomitant loss of enzyme activity. Competitive protection against modification and inactivation was afforded by the presence of Ca2+ and/or micellar concentrations of substrate analogue, lysophosphatidylcholine. Neither modification caused any significant conformational change, as judged from circular dichroic properties. Amino acid analyses and the alignment of peptides from cyanogen bromide and proteolytic cleavage of modified enzyme preparations delineated His-45 as the only residue modified by p-bromophenacyl bromide. However, pyridoxal 5'-phosphate was shown to have reacted not with a single lysine but with four different ones (residues 11, 33, 58 and 111) in such a manner that an overall stoichiometry of one modified lysine residue/molecule enzyme resulted. Apparently, the essential function of lysine could be fulfilled by any one out of these four residues.

Acetophenones↗
PubMed1977

The importance of protein structure and conformation in the preparation of phospholipase-free cardiotoxin from snake venom.

Hydrophobic interactions of cobra venom phospholipase A2 (Mr 13 400) in saline with Sephadex gels and its stability towards denaturation in 6 M guanidine-hydrochloride precluded the use of these solvents to remove traces (approx. 0.2%, w/w) of phospholipase A2 from cardiotoxin (Mr 6800) by gel chromatography. Phospholipase-free (less than 0.001%, w/w) cardiotoxin could, however, be obtained by gel chromatography in 8 M urea or 80 mM phenylalanine. Stokes radius and circular dichroism measurements showed that the hydrophobic retardation of phospholipase A2 was abolished but that the hydrodynamic size and conformation of neither protein was affected, thereby facilitating separation.

Animals↗
PubMed1976

An essential tryptophan in the active site of phospholipase A2 from the venom of Bitis gabonica.

The role of tryptophan in phospholipase A2 (EC 3.1.1.4) from the venom of the gaboon viper, Bitis gabonica, has been investigated. Modification of the enzyme with N-bromosuccinimide and 2-nitrophenylsulfenylchloride showed that the two tryptophan residues in the enzyme, viz. Trp-28 and Trp-59, differ in reactivity towards the reagents. Only Trp-28 reacted with N-bromosuccinimide while a preferential reaction occurred between Trp-59 and 2-nitrophenyl-sulfenylchloride. In each case it was found that loss of enzyme activity was specifically correlated with modification of TRP-28. CD spectra indicated that neither the local nor the gross conformation of the enzyme was altered by modification of Trp-28 and it was therefore concluded that Trp-28 is crucial for enzyme activity. The active enzyme was protected against N-bromosuccinimide inactivation by micellar concentrations of substrate or substrate analogue, suggesting that Trp-28 is involved in substrate binding.

Amino Acid Sequence↗
PubMed1975

A comparison of DNA from free living and endosymbiotic Rhizobium leguminosarum (strain PRE).

1. Bacteroids of Rhizobium leguminosarum (strain PRE) purified from root nodules of Pisum sativum (var. 'Rondo') by the standard procedure of differential centrifugation contained considerable contamination of mitochondrial material. This could be removed by incubation of the bacteroid preparation with 1 M KCl/1% deoxycholate. 2. The DNA content of bacteroid cells of R. leguminosarum was found to have increased about three fold in comparison with the DNA content of free living R. leguminosarum bacteria. 3. No significant difference in DNA composition of free living R. leguminosarum bacteria and bacteroids could be detected by CsCl equilibrium centrifugation, RNA - DNA hybridization and DNA - DNA reassociation studies.

Atypical Bacterial Forms↗
PubMed1975

The two-domain structure of cytochrome b5 in deoxycholate solution.

The membrane protein cytochrome b5 and the polar and hydrophobic fragments into which it is cleaved by trypsin have been investigated, with major emphasis on the deoxycholate-solubilized form of the protein. Molecular weight measurements show that both the intact protein and the fragments are in a monomeric state in deoxycholate and that a small peptide of perhaps 15 residues is excised when the fragments are formed. Measurements of Stokes radius show that the major fragments are globular, but that intact cytochrome b5 has an asymmetric shape, consistent with a structure composed of two globular domains joined by a link region that may be as long as 30 to 40 A. Circular dichroism measurements were made in the far-ultraviolet and in the Soret region, and they add to previously existing data to make it virtually certain that the polar heme-containing domain is unaffected by proteolysis or by removal of deoxycholate. A significant change in the ultraviolet circular dichroism is, however, observed when proteolysis occurs and it is likely that it arises from the link between the domains, which appears to be highly structured (perhaps helical) in the intact protein, but randomly coiled after it is excised. The binding studies reported previously from this laboratory suggest that these inferences about the structure of cytochrome b5 in deoxycholate solution apply also to the protein as solubilized by detergent micelles, by phospholipid vesicles, or by the microsomal membrane.

Animals↗
PubMedDate not supplied

Plasma vitamin levels in patients on prolonged total parenteral nutrition.

Vitamins are essential in total parenteral nutrition (TPN), their importance being highlighted by repeated past documentation of various vitamin deficiencies particularly in patients on long-term parenteral nutrition therapy. This study evaluated the efficacy of water- (Soluvit) and fat-soluble (Vitalipid) vitamin supplementation in patients receiving total parenteral nutrition using the three in one 3-liter bag system. All patients received water-soluble vitamin supplements daily. Fat-soluble vitamin supplements were administered on a daily or twice weekly basis. Twenty-two patients were studied. In seven of the 22 patients vitamin status was assessed on more than one occasion during TPN support, thus bringing the total number of observations to 30. The mean duration of TPN support was 35 days. Eight of the observations were made during less than 10 days, three between 11-19 days, 15 between 20-60 days, and four during more than 60 days of TPN support. Biochemical deficiency as judged by subnormal enzyme activity or vitamin levels were present in 10% of the patients for thiamin, 3% for riboflavin, and 6% for nicotinic acid. By contrast 83% of the patients had low plasma vitamin C and B6 levels. Low plasma vitamin A and E levels were also present in 43 and 40% of the patients, respectively. According to the plasma concentrations of the vitamins studied it would appear that the commercial vitamin preparations used in this study are inadequate in maintaining optimal vitamin status.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMedDate not supplied

Monoclonal antibody characterization of two field strains of Haemophilus paragallinarum isolated from vaccinated layer hens.

An oil-based bacterin, containing strains 083 and 0222 of Haemophilus paragallinarum, is commonly used in South Africa to vaccinate laying flocks against infectious coryza. Two strains of H. paragallinarum, designated M85 and SB86, were isolated from infected but vaccinated commercial laying flocks in two incidental outbreaks of coryza in 1985 and 1986. A panel of five monoclonal antibodies was established which clearly distinguished the vaccine strains from the field isolates. One of these reacted with only vaccine strains A and B, another reacted with only field strains M85 and SB86, and the remaining three cross-reacted to various degrees with all four strains or isolates. Immunoassays were performed by enzyme-linked immunosorbent assay using whole bacteria as solid-phase antigen. These monoclonal antibodies may aid in serotyping new field isolates of H. paragallinarum and in improved standardization of vaccine strains.

Animals↗