Characterization and cDNA cloning of an immune-induced lysozyme from cultured Aedes albopictus mosquito cells

Hernandez, Vida P.; Higgins, LeeAnn; Fallon, Ann M.

doi:10.1016/s0145-305x(02)00065-4

Cited by 13 publications

(8 citation statements)

References 29 publications

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“…An increase in c-type lysozyme gene expression following immune challenge has been reported for several Lepidoptera (Fujimoto et al, 2001; Lee & Brey, 1995; Sun et al, 1991; Mulnix and Dunn, 1994) and for adult mosquitoes (Li et al, 2005) and cultured cells from Aedes aegypti or A. albopictus (Gao & Fallon, 2000; Hernandez et al, 2003). The strong increase in Lys c-1 activity after immune challenge with Gram-negative or Gram-positive bacteria is intriguing since Lys c-1 can directly kill only a small number of Gram-positive bacterial species in vitro .…”

Section: Discussionmentioning

confidence: 91%

Characterization of expression, activity and role in antibacterial immunity of Anopheles gambiae lysozyme c-1

Kajla

Andreeva

Gilbreath

et al. 2010

Comparative Biochemistry and Physiology Part B: Biochemistry an

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There are eight lysozyme genes in the Anopheles gambiae genome. Transcripts of one of these genes, LYSC-1, increased in Anopheles gambiae cell line 4a3B by 24 h after exposure to heat-killed Micrococcus luteus. Lysozyme activity was also identified in conditioned media from the cell line from which the protein was purified to homogeneity using ion exchange and gel filtration. Mass spectrometric analysis of the purified protein showed 100% identity to lysozyme c-1. Purified lysozyme c-1 was tested against non-mosquito derived as well as culturable bacteria isolated from mosquito midguts. Lysozyme c-1 had negligible effects on the growth of most mosquito-derived bacteria in vitro but did inhibit the growth of M. luteus. Although Lys c-1 did not directly kill most bacteria, knockdown of LYSC-1 resulted in significant mortality in mosquitoes subjected to hemocoelic infections with Escherichia coli but not M. luteus thus suggesting that this protein plays an important role in antibacterial defense against selected bacteria.

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Section: Discussionmentioning

confidence: 91%

Characterization of expression, activity and role in antibacterial immunity of Anopheles gambiae lysozyme c-1

Kajla

Andreeva

Gilbreath

et al. 2010

Comparative Biochemistry and Physiology Part B: Biochemistry an

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“…A number of studies have shown an increased lysozyme expression in the mosquitoes after challenge with bacteria (Fujimoto et al, 2001; Lee and Brey, 1995; Sun et al, 1991; Gao and Fallon, 2000; Hernandez et al, 2003; Bedoya et al, 2005; Kajla et al, 2010). In blood fed mosquitoes the bacterial population increases significantly within the midgut (Beier, 1998, DeMaio et al, 1996; Pumpuni et al, 1993, 1996; Bedoya et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Investigations on the role of a lysozyme from the malaria vector Anopheles dirus during malaria parasite development

Lapcharoen

Komalamisra

Rongsriyam

et al. 2012

Developmental & Comparative Immunology

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A cDNA encoding a lysozyme was obtained by rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) from females of the malaria vector Anopheles dirus A (Diptera: Culicidae). The 623 bp lysozyme (AdLys c-1) cDNA encodes the 120 amino acid mature protein with a predicted molecular mass of 13.4 kDa and theoretical pI of 8.45. Six cysteine residues and a potential calcium binding motif that are present in AdLys c-1 are highly conserved relative to those of c-type lysozymes found in other insects. RT-PCR analysis of the AdLys c-1 transcript revealed its presence at high levels in the salivary glands both in larval and adult stages and in the larval caecum. dsRNA mediated gene knockdown experiments were conducted to examine the potential role of this lysozyme during Plasmodium berghei infection. Silencing of AdLys c-1 resulted in a significant reduction in the number of oocysts as compared to control dsGFP injected mosquitoes.

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“…The precolumn was switched in-line with a capillary column, and peptides were eluted at 350 nl per min using an LCP Ultimate LC system. The capillary column (100-m internal diameter) was packed in-house to 12 cm in length with 200-Å, 5-m-pore-size C 18 particles (Michrom BioResources, Auburn, CA) as described previously by Hernandez et al (11). Peptides were eluted from the column and introduced into a QSTAR Pulsar quadrupole-TOF MS which was equipped with Protana's nano-electrospray source under the same conditions described previously by Kapphahn et al (13) Louis, MO) were used for external calibration.…”

Section: Methodsmentioning

confidence: 99%

Purification and Characterization of Allophanate Hydrolase (AtzF) from Pseudomonas sp. Strain ADP

2005

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AtzF, allophanate hydrolase, is a recently discovered member of the amidase signature family that catalyzes the terminal reaction during metabolism of s-triazine ring compounds by bacteria. In the present study, the atzF gene from Pseudomonas sp. strain ADP was cloned and expressed as a His-tagged protein, and the protein was purified and characterized. AtzF had a deduced subunit molecular mass of 66,223, based on the gene sequence, and an estimated holoenzyme molecular mass of 260,000. and hydroxylamine to generate malonohydroxamate, potentially derived from hydroxylamine capture of an enzyme-tethered acyl group. Three putative catalytically important residues, one lysine and two serines, were altered by site-directed mutagenesis, each with complete loss of enzyme activity. The identity of a putative serine nucleophile was probed using phenyl phosphorodiamidate that was shown to be a time-dependent inhibitor of AtzF. Inhibition was due to phosphoroamidation of Ser189 as shown by liquid chromatography/matrix-assisted laser desorption ionization mass spectrometry. The modified residue corresponds in sequence alignments to the nucleophilic serine previously identified in other members of the amidase signature family. Thus, AtzF affects the cleavage of three carbon-to-nitrogen bonds via a mechanism similar to that of enzymes catalyzing single-amide-bond cleavage reactions. AtzF orthologs appear to be widespread among bacteria.

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Characterization and cDNA cloning of an immune-induced lysozyme from cultured Aedes albopictus mosquito cells

Cited by 13 publications

References 29 publications

Characterization of expression, activity and role in antibacterial immunity of Anopheles gambiae lysozyme c-1

Characterization of expression, activity and role in antibacterial immunity of Anopheles gambiae lysozyme c-1

Investigations on the role of a lysozyme from the malaria vector Anopheles dirus during malaria parasite development

Purification and Characterization of Allophanate Hydrolase (AtzF) from Pseudomonas sp. Strain ADP

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