Amino Acid Sequence Of Lysozyme Research Articles

Epitope Mapping of Lysozyme Using the Chinese Egg-Allergic Sera at Both Pooled and Individual Levels

To accurately map the B-cell linear epitopes of lysozyme (LYS) in eggs, five bioinformatics tools were first used to obtain the mimotopes. Afterward, based on the Chinese egg-allergic sera samples screened by the indirect enzyme-linked immunosorbent, the epitopes possessing the capability of binding to IgG/IgE were mapped at both pooled and individual levels by using overlapping peptides covering the complete amino acid sequence of LYS. Six B-cell linear epitopes and two dominant B-cell linear epitopes that could bind to LYS-sIgG were mapped for the first time. Seven IgE-binding epitopes and three dominant IgE-binding epitopes were also obtained. Furthermore, AA31-34 and AA88-91 were the shared dominant epitopes of LYS-sIgG and LYS-sIgE at pooled and individual levels. Overall, the mapped B-cell linear epitopes filled in the gaps in the study of LYS epitopes, and the results may provide theoretical support for the following immunotherapy of egg allergy.

CDNA cloning of the lysozyme of the white shrimp Penaeus vannamei

Lysozyme, an antibacterial protein, has been implicated in innate immunity in invertebrates, but its activity in shrimp remained to be determined. We cloned the white shrimp lysozyme cDNA using a PCR strategy and detected its activity in haemocytes using a lytic-zone assay against Micrococcus luteus. The cloning was based on a reported EST (dbEST BE18831). The deduced amino acid sequence resulted in 150 amino with 46% identity to hen egg white lysozyme. RT-PCR was used to detect lysozyme mRNA in haemocytes. Analysis of the amino acid sequence of the shrimp lysozyme showed that it belongs to the C-type family of lysozymes. Furthermore, the lysozyme amino acid sequence contained extra residues at its C-terminus, which are characteristic of marine invertebrates. This information will be useful in future studies on the molecular mechanisms of immunity in marine invertebrates.

Amino acid sequences of hemoglobin from guinea fowl (Numida meleagri) and California quail (Lophortyx californica) with phylogenetic analysis of major groups of Galliformes.

We determined the complete amino acid sequences of the hemoglobin of two species, guinea fowl and California quail, in Galliformes from intact globin chain and chemical cleavage fragments in order to analyze the molecular evolution of hemoglobin for the classification of Galliformes. Galliformes have two types of hemoglobin components, HbA and HbD, which consist of identical beta chain and different alpha chains. The sequences are similar to globin chains of Galliformes reported previously. These sequences were compared with those of other Galliformes (Phasianidae, Meleagrididae) using duck and goshawk as out-groups. The phylogenetic tree of major groups of Galliformes based on hemoglobin was similar to the tree model produced based on the amino acid sequence of lysozyme c.

Larval and pupal induction and N-terminal amino acid sequence of lysozyme from Heliothis virescens

Fifth instar larvae and prepupae of Heliothis virescens (tobacco budworm) were injected with live Enterobacter cloacae and bled at different times after vaccination. Immune pupal hemolymph showed a 54 times increase in lysozyme activity when compared with normal larval hemolymph, and an 11 times increase of lysozyme activity when compared with immune larval hemolymph. Lysozyme activity of the normal pupal hemolymph increased as greatly as did lysozyme activity of the immune larval hemolymph after metamorphosis. The pupal immune response with regard to lysozyme was much greater than the larval immune response in H. virescens. Lysozyme was purified by heat treatment at 100°C and a chromatography series that included reverse-phase HPLC. The molecular mass of H. virescens lysozyme was approximately 16 kDa by SDS–PAGE which is greater than other insect lysozymes and chicken lysozyme. Amino acid sequence of the N-terminus showed that H. virescens lysozyme is 82% homologous with lysozyme of Manduca sexta and Galleria mellonella. CNBr cleavage of H. virescens lysozyme produced 11 and 6 kDa peptide fragments indicating that one methionine was present, which was also supported by amino acid analysis. However, methionine was located at the carboxyl terminal side rather than the N-terminal side as judged by the N-terminal sequences of each peptide fragment. The residue 22 in most lepidopteran lysozymes is methionine, whereas H. virescens lysozyme had a leucine at residue 22. There was an amino acid deletion near the carboxyl terminal side of H. virescens lysozyme as also found in Trichoplusia ni.

One step isolation of human lysozyme using exclusion chromatography based on isoelectric point

Lysozyme (mucopeptide N-acetylmuramoylhydrolase EC 3.2.1.17) has been extensively studied during the last decades. This protein, of low molecular mass and high isoelectric point, well known for its lytic activity, is found in many sources such as organs of vertebrates, egg white, plants, bacteriophage, milk, tears, etc. Data from Jolles and Jolles [l] and Osserman and Lawlor [2] indicate the probable identity of the enzyme in all human tissues and secretions. Furthermore, the amino acid sequence of a lysozyme extracted from urine of a patient with leukemia has been found to be exactly the same as the amino acid sequence of human milk lysozyme [3]. But human lysozymes show several significant differences in their composition and also in their structure as compared to bird egg white lysozymes. We have been interested in the development of techniques for the isolation and analysis of enzymes and biological macromolecules which could permit the use of extremely small quantities of starting material [4]. One of these problems was the isolation in pure form of lysozymes from samples of human tears [5] for utilisation in clinical assays. Until now the purification of lysozymes has been accomplished in different ways usually involving several steps: among them precipitation at pH 9.5 [6], ion exchange and gel filtration [1,7], affinity chromatography [8] or interaction with agarose [9]. In this paper we describe a rapid one-step chromatographic procedure for the purification of lacrymal and urinary lysozymes. This purified enzyme can be used as standard for lacrymal lysozyme assay for diagnosis of Sjiigren syndrome.

Amino acid sequence and immunological properties of chalchalaca egg white lysozyme.

The amino acid sequence of lysozyme c from chachalaca egg white was determined. Like other bird lysozymes c, that of the chachalaca has 129 amino acid residues. It differs from other avian lysozymes c by 2 to 31 amino acid substitutions as well as by being devoid of phenylalanine. It contains substitutions at 9 positions which are invariant in the other 7 bird lysozymes of known sequence. Although the chachalaca is classified zoologically in the order Galliformes, which includes chickens and other pheasant-like birds, its lysozyme differs more from those of pheasant-like birds than do the lysozymes c of ducks. Phylogenetic analysis of the sequence comparisons confirms that the lineage leading to chachalaca lysozyme c separated from that leading to other galliform lysozymes c before the duck lysozyme c lineage did. This indicates a contrast between protein evolution and evolution at the organismal level. Immunological comparison of chachalaca lysozyme c with other lysozymes of known sequence provides further support for the proposal that immunological cross-reactivity is strongly dependent on degree of sequence resemblance among bird lysozymes.

Amino acid sequence of lysozyme from baboon milk

Reduced alkylated baboon milk lysozyme was subjected to digestion with trypsin. The resulting peptides were purified by a combination of Dowex 1 (X2) and chromatography and electrophoresis on paper. The amino acid sequence of these peptides was determined in detail chiefly by the Edman procedure. Alignment of the tryptic peptides into a single chain containing 130 amino acids was established chiefly by homology with human milk lysozyme; 14 replacements were noted between the two enzymes. Baboon milk lysozyme was devoid of methionine and contained six basic amino acids (arginine residues) less than human milk lysozyme.

X-ray crystallography and the amino acid sequence of lysozyme.

Certain parts of the amino acid sequence of hen's-egg lysozyme were reinvestigated. The parts in question are those in which X-ray-crystallographic data were used to decide between the two reported versions of the sequence that were based on chemical data. We conclude that the X-ray evidence, in the main, led to the correct choice between the sequences.

RECENT DEVELOPMENTS IN THE STUDY OF LYSOZYMES.

AbstractLysozymes are widely distributed enzymes. They split the chemical skeleton of bacterial cell walls by hydrolysing the β (1 → 4) linkages between N‐acetylmuramic acid and N‐acetylglucosamine. the amino acid sequence of lysozyme from hen's egg‐white is known (129 amino acid residues). Lysozymes of different origins frequently differ in their primary structures although they exhibit qualitatively the same biological activity. The results obtained from investigations with lysozymes allow conclusions to be drawn regarding the chemical structure of the “backbone” of bacterial cell walls.