Properties of Drosophila melanogaster prophenoloxidases expressed in Escherichia coli

Li, Xuquan; Ma, Mingliang; Liu, Fei; Yang, Chen; Lü, Aijun; Ling, Qing; Li, Jianyong; Beerntsen, Brenda T.; Yu, Xiao‐Qiang; Liu, Chao-Liang; Ling, Erjun

doi:10.1016/j.dci.2011.11.005

Cited by 45 publications

(54 citation statements)

References 26 publications

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“…By contrast, only DmPPO1 and DmPPO2 were expressed in wild-type D. melanogaster (Tang, 2009; Binggeli et al, 2014). Another study found that recombinant DmPPOs were expressed in E. coli at 16°C, yielding soluble DmPPO (Li et al, 2012). These recombinant DmPPO proteins showed PO activity upon activation by cetylpyridinium chloride (CPC), ethanol, and serine protease.…”

Section: Expression Of Recombinant Ppo In Eukaryotic and Prokaryotic mentioning

confidence: 99%

Insect prophenoloxidase: the view beyond immunity

et al. 2014

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Insect prophenoloxidase (PPO) is an important innate immunity protein due to its involvement in cellular and humoral defense. It belongs to a group of type-3 copper-containing proteins that occurs in almost all organisms. Insect PPO has been studied for over a century, and the PPO activation cascade is becoming clearer. The insect PPO activation pathway incorporates several important proteins, including pattern-recognition receptors (PGRP, β GRP, and C-type lectins), serine proteases, and serine protease inhibitors (serpins). Due to their complexity, PPO activation mechanisms vary among insect species. Activated phenoloxidase (PO) oxidizes phenolic molecules to produce melanin around invading pathogens and wounds. The crystal structure of Manduca sexta PPO shows that a conserved amino acid, phenylalanine (F), can block the active site pocket. During activation, this blocker must be dislodged or even cleaved at the N-terminal sequence to expose the active site pockets and allow substrates to enter. Thanks to the crystal structure of M. sexta PPO, some domains and specific amino acids that affect PPO activities have been identified. Further studies of the relationship between PPO structure and enzyme activities will provide an opportunity to examine other type-3 copper proteins, and trace when and why their various physiological functions evolved. Recent researches show that insect PPO has a relationship with neuron activity, longevity, feces melanization (phytophagous insects) and development, which suggests that it is time for us to look back on insect PPO beyond the view of immunity in this review.

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Section: Expression Of Recombinant Ppo In Eukaryotic and Prokaryotic mentioning

confidence: 99%

Insect prophenoloxidase: the view beyond immunity

et al. 2014

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“…After incubation at room temperature for 30 min, 0.8 mg of Drosophila PPO1 was added to the reaction mixtures, and incubated at 37 C for 1 h. The mixtures were treated with 5 Â SDS sample buffer and subjected to immunoblot analysis using antiserum against Drosophila PPO1 (1:2000 diluted) (Li et al, 2012) or His tag (1:1,1000 diluted).…”

Section: Analysis Of Sp13 Inhibition Using Recombinant Propo As Substmentioning

confidence: 99%

“…In mutant proSP1 (proSP1 Xa ), the cleavage activation site of proSP1 was changed from LDLH 114 to IEGR 114 to permit its activation by bovine Factor Xa. Recombinant Drosophila PPO1 was produced and purified as described by Li et al (Li et al, 2012).…”

Section: Production Of Recombinant Proteins and Preparation Of Antisementioning

confidence: 99%

Ostrinia furnacalis serpin-3 regulates melanization cascade by inhibiting a prophenoloxidase-activating protease

Cheng

Zhou

Liu

et al. 2015

Insect Biochemistry and Molecular Biology

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“…The activation of recombinant prophenol oxidase (rPPO) in vitro was performed as previously reported, with necessary changes (Li et al, 2012a). CDEP-1 (100 ng) and rPPO1eHis (300 ng) were preincubated for 1 h at 4 C, then mixed with 200 ml of dopamine solution, and incubated for 5 min at room temperature.…”

Section: Phenol Oxidase Activationmentioning

confidence: 99%