Abstract:
:The second domain (R-020) of human urinary trypsin inhibitor (UTI) exerts similar inhibitory activities on trypsin, alpha-chymotrypsin, leukocyte elastase, and plasmin to those of UTI itself, and additionally inhibits coagulation factor Xa (FXa) and plasma kallikrein, on both of which UTI has no inhibitory effect. In the present study, we attempted to increase this FXa-inhibitory activity by modeling the structure of R-020-FXa complex and substituting one or two amino acids in R-020 using recombinant DNA technology. Molecular modeling of R-020 and FXa was performed with reference to X-ray analysis of the complex of bovine pancreatic trypsin inhibitor (BPTI) and bovine trypsin to determine the site of amino acid modification. The expression plasmids into which R-020 genes with base substitution were inserted were prepared and introduced into Escherichia coli to express R-020 variants. The resulting variants were purified and their enzyme inhibitory activities were measured. The FXa-inhibitory activity was increased in four variants with single amino acid substitution. With another four variants having two amino acid substitutions involving combinations of the above single amino acid substitutions, the FXa-inhibitory activity was further increased. Because the electrostatic interaction within R-020-FXa complex seemed stronger in these R-020 variants, this increase in FXa-inhibitory effect was speculated to be a consequence of more potent binding between the enzyme and the inhibitor.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Nii A,Morishita H,Yamakawa T,Matsusue T,Hirose J,Miura T,Isaji M,Horisawa Y,Sugihara K,Kanamori Tdoi
10.1093/oxfordjournals.jbchem.a124465subject
Has Abstract,Author List Incompletepub_date
1994-06-01 00:00:00pages
1107-12issue
6eissn
0021-924Xissn
1756-2651journal_volume
115pub_type
杂志文章abstract::Pure recA protein was easily obtained from Escherichia coli harboring plasmid pTM-2 which carried the recA gene by two chromatographic steps on phosphocellulose and DEAE-cellulose. RecA protein was stable in the pH range of 6 to 9 at 25 degrees C. RecA protein was found to aggregate highly under these conditions. Lowe...
journal_title:Journal of biochemistry
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更新日期:1981-10-01 00:00:00
abstract::Rat brain type II (beta) protein kinase C (PKC) was phosphorylated by rat lung casein kinase II (CK-II). Neither type I (gamma) nor type III (alpha) PKC was significantly phosphorylated by CK-II. CK-II incorporated 0.2-0.3 mol of phosphate into 1 mol of type II PKC. This phosphate was located at the single seryl resid...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123635
更新日期:1991-10-01 00:00:00
abstract::We purified an antimicrobial protein of 76 residues, denoted bovine antimicrobial protein-1 (BAMP-1), from fetal calf serum using hydrophobic chromatography, gel filtration, and reverse-phase high-performance liquid chromatography. The amino acid sequence of BAMP-1 was similar to that of human apolipoprotein A-II (apo...
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更新日期:1998-04-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134719
更新日期:1984-04-01 00:00:00
abstract::Guanidine hydrochloride (GdnHCl)-induced unfolding of bovine spleen galectin-1 (Gal-1) exhibits three-state mechanism involving exclusive, structured tertiary monomer in 0.5 M GdnHCl. Gal-1 has one tryptophan residue (Trp 68) per subunit. Phosphorescence spectra of both Gal-1 dimer and intermediate monomer at 77 K sho...
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abstract::Antizyme (AZ) interacts with ornithine decarboxylase, which catalyzes the first step of polyamine biosynthesis and recruits it to the proteasome for degradation. Synthesizing the functional AZ protein requires transition of the reading frame at the termination codon. This programmed +1 ribosomal frameshifting is induc...
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abstract::The role of thiols on the activation and/or stabilization of rat brain nitric oxide synthase (NOS) has been investigated. It was found that thiols are not necessary for stabilizing or protecting the protein during purification but are required during enzyme turnover for maximum activity. In the complete absence of thi...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124797
更新日期:1995-04-01 00:00:00
abstract::Four motor proteins, MotX, MotY, PomA, and PomB, have been identified as constituents of the Na(+)-driven flagellum of Vibrio species. In this study, the complete motX gene was cloned from Vibrio alginolyticus and shown to complement three mot mutations, motX94, motX115, and motX119, as well as a V. parahaemolyticus m...
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132723
更新日期:1980-01-01 00:00:00
abstract::Amyloid formation is the hallmark of many diseases. The propensity of a protein to aggregate depends on a number of biological factors like the presence of sulphated polysaccharides termed as glycosaminoglycans (GAGs). Here we assessed whether the polymeric nature of GAGs is responsible for their protein aggregation-p...
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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journal_title:Journal of biochemistry
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更新日期:2005-10-01 00:00:00
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a131253
更新日期:1976-07-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134740
更新日期:1984-05-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a122440
更新日期:1988-08-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134682
更新日期:1984-03-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1983-11-01 00:00:00
abstract::The gizzard tropomyosin molecule is composed of two subunits at 1:1 molar ratio. Possible composites of the tropomyosin molecule are two kinds of homodimer (one for each subunit), a heterodimer of two subunits, or a mixture of heterodimer and homodimer(s). We tried to evaluate the native subunit composition of gizzard...
journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1991-05-01 00:00:00
abstract::The technique of saturation transfer electron spin resonance has been applied to study the rotational diffusion of spin-labeled Ca2+, Mg2+-dependent ATPase molecules in the membranes of sarcoplasmic reticulum vesicles. Comparison of the present data with those for spin-labeled hemoglobin undergoing isotropic rotation ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132099
更新日期:1978-07-01 00:00:00
abstract::3-Isopropylmalate dehydrogenase (IPMDH) is a dimeric enzyme with a strongly hydrophobic core that is composed of residues from four alpha-helices. We replaced Glu253, which is found in the hydrophobic core and is part of the subunit interface of the Bacillus subtilis (Bs) IPMDH, with several other amino acids to probe...
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更新日期:1990-10-01 00:00:00
abstract::Prp8 is a splicing factor of 220 kDa originally identified in yeast and is a component of the U5 small nuclear ribonucleoprotein particle. Mouse Prp8 cDNA was cloned and shown to share 62.6 and 68.2% sequence identity with the yeast homologue at the amino acid and nucleotide level, respectively, while it differs by on...
journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:2001-04-01 00:00:00
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journal_title:Journal of biochemistry
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更新日期:1995-05-01 00:00:00
abstract::Temperature and solvent effects on reaction center structures were examined in two thermophilic photosynthetic bacteria, Chloroflexus aurantiacus and Chromatium tepidum, in order to gain insight into the interactions among the reaction center proteins and pigment systems. Thermal stability of the reaction centers was ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1991-10-01 00:00:00