Application of methyl parathion hydrolase (MPH) as a labeling enzyme

Wei Yang; Ya-Feng Zhou; He-Ping Dai; Li-Jun Bi; Zhi-Ping Zhang; Xiao-Hua Zhang; Yan Leng; Xian-En Zhang
April 2008
Analytical & Bioanalytical Chemistry;Apr2008, Vol. 390 Issue 8, p2133
Academic Journal
Methyl parathion hydrolase (MPH) is an enzyme that catalyzes the degradation of methyl parathion, generating a yellow product with specific absorption at 405 nm. The application of MPH as a new labeling enzyme was illustrated in this study. The key advantages of using MPH as a labeling enzyme are as follows: (1) unlike alkaline phosphatase (AP), horseradish peroxidase (HRP), and glucose oxidase (GOD), MPH is rarely found in animal cells, and it therefore produces less background noise; (2) its active form in solution is the monomer, with a molecular weight of 37 kDa; (3) its turnover number is 114.70 ± 13.19 s−1, which is sufficiently high to yield a significant signal for sensitive detection; and (4) its 3D structure is known and its C-terminal that is exposed to the surface can be easily subjected to the construction of genetic engineering monocloning antibody–enzyme fusion for enzyme-linked immunosorbent assay (ELISA). To demonstrate its utility, MPH was ligated to an single-chain variable fragment (scFv), known as A1E, against a white spot syndrome virus (WSSV) with the insertion of a [–(Gly–Ser)5–] linker peptide. The resulting fusion protein MPH-A1E possessed both the binding specificity of the scFv segment and the catalytic activity of the MPH segment. When MPH-A1E was used as an ELISA reagent, 25 ng purified WSSV was detected; this was similar to the detection sensitivity obtained using A1E scFv and the HRP/Anti-E Tag Conjugate protocol. The fusion protein also recognized the WSSV in 1 μL hemolymph from an infected shrimp and differentiated it from a healthy shrimp. [Figure not available: see fulltext.]


Related Articles

  • Critical Active-Site Residues Identified by Site-Directed Mutagenesis in Pseudomonas aeruginosa Phosphorylcholine Phosphatase, A New Member of the Haloacid Dehalogenases Hydrolase Superfamily. Beassoni, Paola R.; Otero, Lisandro H.; Massimelli, Maria J.; Lisa, Angela T.; Domenech, Carlos E. // Current Microbiology;Dec2006, Vol. 53 Issue 6, p534 

    Pseudomonas aeruginosa phosphorylcholine phosphatase (PChP), the product of the PA5292 gene, is synthesized when the bacteria are grown with choline, betaine, dimethylglycine, or carnitine. In the presence of Mg2+, PChP catalyzes the hydrolysis of both phosphorylcholine (PCh) and...

  • Phenol oxidase is a necessary enzyme for the silkworm molting which is regulated by molting hormone. Wang, Mei-xian; Lu, Yan; Cai, Zi-zheng; Liang, Shuang; Niu, Yan-shan; Miao, Yun-gen // Molecular Biology Reports;May2013, Vol. 40 Issue 5, p3549 

    Insect molting is an important developmental process of metamorphosis, which is initiated by molting hormone. The molting process includes the activation of dermal cells, epidermal cells separation, molting fluid secretion, the formation of new epidermis and old epidermis excoriation etc....

  • HIV-1 Protease Inhibitor : A Review. Toraskar, Mrunmayee P.; Dhanashire, Shweta T.; Singh, Pratima R. P.; Kadam, Vilasrao J. // Journal of Pharmacy Research;Dec2009, Vol. 2 Issue 12, p1818 

    HIV-1 is notoriously lethal virus that is known to cause AIDS. This HIV virus contains RNA that is translated into a polypeptide sequence, that sequence is assembled in a long chain that includes several individual proteins (reverse transcriptase, protease, integrase). Before these enzymes...

  • In this issue.  // Nature Reviews Drug Discovery;Jan2012, Vol. 11 Issue 1, p1 

    An introduction is presented in which the editor discusses various reports within the issue on topics including the advances in computer-assisted peptide design strategies in the creation of antimicrobial peptides (AMPs), the concept of interfacial inhibition, and the therapeutic use of...

  • Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei. Song, Xiaoling; Zhang, Yue; Wei, Song; Huang, Jie // Chinese Journal of Oceanology & Limnology;Mar2013, Vol. 31 Issue 2, p374 

    The effects of different hydrolysis methods on peptidoglycan (PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp, Litop enaeus vannamei. PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with...

  • ATP generation in a host cell in early-phase infection is increased by upregulation of cytochrome c oxidase activity via the p2 peptide from human immunodeficiency virus type 1 Gag. Minako Ogawa; Yuki Takemoto; Shintaro Sumi; Daisuke Inoue; Naoki Kishimoto; Nobutoki Takamune; Shozo Shoji; Shinya Suzu; Shogo Misumi // Retrovirology;11/17/2015, Vol. 12, p1 

    Background: Human immunodeficiency virus type 1 (HIV-1) must take advantage of its own proteins with two or more functions to successfully replicate. Although many attempts have been made to determine the function of viral proteins encoded in the HIV-1 genome, the role of the p2 peptide, a...

  • Expression and Functional Studies of Ubiquitin C-Terminal Hydrolase L1 Regulated Genes. Bheda, Anjali; Shackelford, Julia; Pagano, Joseph S. // PLoS ONE;2009, Vol. 4 Issue 8, p1 

    Deubiquitinating enzymes (DUBs) have been increasingly implicated in regulation of cellular processes, but a functional role for Ubiquitin C-terminal Hydrolases (UCHs), which has been largely relegated to processing of small ubiquitinated peptides, remains unexplored. One member of the UCH...

  • Peptide inhibitors of HIV-1 and HIV-2 proteases: a comparative study. PICHOVÁ, Iva; WEBER, Jan; LITERA, Jaroslav; KONVALINKA, Jan; VONDRÁSEK, JiřI; SOUČEK, Milan; STROP, Petr; MAJER, Pavel; HEUSER, Anke M.; KRAEUSSLICH, Hans-Georg // Leukemia (08876924);Apr97 Supplement 3, Vol. 11, p120 

    HIV-1 and HIV-2 proteases (PR) which play the key role in the formation of infectious viral particles offer a target for inhibitors that could block the maturation step. Inhibitors o HIV-1 PR exhibit mostly 1 - 2 orders of magnitude weaker affinity lot HIV-2 PR. The subsite specificity study of...

  • Hormonal regulation of energy metabolism in insects as a driving force for performance. Lorenz, Matthias W.; Gäde, Gerd // Integrative & Comparative Biology;Oct2009, Vol. 49 Issue 4, p380 

    Since all life processes depend on energy, the endocrine control of energy metabolism is one of the driving forces for the performance of an individual. Here, we review the literature on the key players in the endocrine regulation of energy homeostasis in insects, the adipokinetic hormones....


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics