High-level expression and characterization of two serine protease inhibitors from Trichinella spiralis

Zhaoxia Zhang; Yixian Mao; Da Li; Yvhan Zhang; Wei Li; Honglin Jia; Jun Zheng; Li Li; Yixin Lu

doi:10.1016/j.vetpar.2016.02.003

High-level expression and characterization of two serine protease inhibitors from Trichinella spiralis

Vet Parasitol. 2016 Mar 30:219:34-9. doi: 10.1016/j.vetpar.2016.02.003. Epub 2016 Feb 3.

Authors

Zhaoxia Zhang¹, Yixian Mao¹, Da Li¹, Yvhan Zhang¹, Wei Li¹, Honglin Jia², Jun Zheng², Li Li¹, Yixin Lu³

Affiliations

¹ Key Laboratory of Animal Common Disease Prevention, College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Harbin 150030, PR China.
² State Key Laboratory of Veterinary Biotechnology-Michigan State University Joint Laboratory of Innate Immunity, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Maduan Street 427, Nangang District, Harbin 150001, PR China.
³ Key Laboratory of Animal Common Disease Prevention, College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Harbin 150030, PR China. Electronic address: luyixin@neau.edu.cn.

PMID: 26921036
DOI: 10.1016/j.vetpar.2016.02.003

Abstract

Serine protease inhibitors (SPIs) play important roles in tissue homeostasis, cell survival, development, and host defense. So far, SPIs have been identified from various organisms, such as animals, plants, bacteria, poxviruses, and parasites. In this study, two SPIs (Tsp03044 and TspAd5) were identified from the genome of Trichinella spiralis and expressed in Escherichia coli. Sequence analysis revealed that these two SPIs contained essential structural motifs, which were well conserved within the tumor-infiltrating lymphocytes (TIL) and serpin superfamily. Based on protease inhibition assays, the recombinant Tsp03044 showed inhibitory effects on trypsin, α-chymotrypsin, and pepsin, while the recombinant TspAd5 could effectively inhibit the activities of α-chymotrypsin and pepsin. Both these inhibitors showed activity between 28 and 48 °C. The expression levels of the two SPIs were also determined at different developmental stages of the parasite with real-time PCR. Our results indicate that Tsp03044 and TspAd5 are functional serine protease inhibitors.

Keywords: Chymotrypsin; Pepsin; Serine protease inhibitor; Trichinella spiralis; Trypsin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Enzyme Activation / drug effects
Enzymes / metabolism
Escherichia coli / genetics
Female
Gene Expression Profiling
Gene Expression Regulation*
Helminth Proteins / chemistry
Helminth Proteins / genetics*
Helminth Proteins / isolation & purification
Helminth Proteins / pharmacology*
Host-Parasite Interactions
Mice
Molecular Sequence Data
Phylogeny
Recombinant Proteins / genetics
Sequence Alignment
Serine Proteinase Inhibitors / genetics*
Serine Proteinase Inhibitors / isolation & purification
Serine Proteinase Inhibitors / pharmacology*
Serpins / genetics
Serpins / isolation & purification
Serpins / pharmacology
Temperature
Trichinella spiralis / classification
Trichinella spiralis / genetics*
Trichinella spiralis / metabolism*

Substances

Enzymes
Helminth Proteins
Recombinant Proteins
Serine Proteinase Inhibitors
Serpins