PMID: 8580909Dec 1, 1995Paper

Expression and characterization of recombinant Manduca sexta serpin-1B and site-directed mutants that change its inhibitory selectivity

Insect Biochemistry and Molecular Biology
Haobo JiangMichael R Kanost

Abstract

Hemolymph of Manduca sexta contains a number of serine proteinase inhibitors from the serpin superfamily. During formation of a stable complex between a serpin and a serine proteinase, the enzyme cleaves a specific peptide bond in an exposed loop (the reactive-site region) at the surface of the serpin. The amino acid residue on the amino-terminal side of this scissile bond, the P1 residue, is important in defining the selectivity of a serpin for inhibiting different types of serine proteinases. M. sexta serpin-1B, with alanine at the position predicted from sequence alignments to be the P1 residue, was previously named alaserpin. This alanyl residue was changed by site-directed mutagenesis to lysine (A343K) and phenylalanine (A343F). The serpin-1B cDNA and its mutants were inserted into an expression vector, H6pQE-60, and the serpin proteins were expressed in Escherichia coli. Affinity-purified recombinant serpins selectively inhibited mammalian serine proteinases: serpin-1B inhibited elastase; serpin-1B(A343K) inhibited trypsin, plasmin, and thrombin; serpin-1B(A343F) inhibited chymotrypsin as well as trypsin. All three serpins inhibited human cathepsin G. This insect serpin and its site-directed mutants associated with mammal...Continue Reading

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Citations

Oct 19, 2010·Cellular and Molecular Life Sciences : CMLS·Chunju AnKristin Michel
Dec 7, 2007·Functional & Integrative Genomics·Thomas H Roberts, Jørn Hejgaard
Jul 30, 1999·Developmental and Comparative Immunology·M R Kanost
Jun 2, 2009·Insect Biochemistry and Molecular Biology·Prashant S PyatiJohn A Gatehouse
Jun 18, 2004·Immunological Reviews·Michael R KanostXiao-Qiang Yu
Sep 8, 2016·Seminars in Cell & Developmental Biology·David A MeekinsKristin Michel
Mar 18, 2004·Archives of Insect Biochemistry and Physiology·K S BrandtN Wisnewski
Jun 20, 2019·Journal of Biomolecular Structure & Dynamics·Prapasiri PongprayoonSathaporn Jittapalapong
May 1, 2004·Biochemistry and Molecular Biology Education : a Bimonthly Publication of the International Union of Biochemistry and Molecular Biology·Joshua A BornhorstAmy B Mulnix
Jun 16, 1999·Structure·J LiE J Goldsmith
Sep 21, 2010·Biochimie·David GubbAne Fullaondo

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