Abstract:
:Trypsin and chymotrypsin have specificity pockets of essentially the same geometry, yet trypsin is specific for basic while chymotrypsin for bulky hydrophobic residues at the P1 site of the substrate. A model by Steitz, Henderson and Blow suggested the presence of a negative charge at site 189 as the major specificity determinant: Asp189 results in tryptic, while the lack of it chymotryptic specificity. However, recent mutagenesis studies have shown that a successful conversion of the specificity of trypsin to that of chymotrypsin requires the substitution of amino acids at sites 138, 172 and at thirteen other positions in two surface loops, that do not directly contact the substrate. For further testing the significance of these sites in substrate discrimination in trypsin and chymotrypsin, we tried to change the chymotrypsin specificity to trypsin-like specificity by introducing reverse substitutions in rat chymotrypsin. We report here that the specificity conversion is poor: the Ser189Asp mutation reduced the activity but the specificity remained chymotrypsin-like; on further substitutions the activity decreased further on both tryptic and chymotryptic substrates and the specificity was lost or became slightly trypsin-like. Our results indicate that in addition to structural elements already studied, further (chymotrypsin) specific sites have to be mutated to accomplish a chymotrypsin-->trypsin specificity conversion.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Venekei I,Szilágyi L,Gráf L,Rutter WJsubject
Has Abstractpub_date
1996-01-29 00:00:00pages
143-7issue
2eissn
0014-5793issn
1873-3468pii
0014-5793(95)01484-5journal_volume
379pub_type
杂志文章相关文献
FEBS LETTERS文献大全abstract::Osteoblasts and adipocytes are believed to share a common progenitor. Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a key role in the switching of these two cell lineages. Here, we demonstrated the differentiation of ES cells into an osteoblastic lineage using siRNA against PPARgamma without the a...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2006.06.057
更新日期:2006-07-24 00:00:00
abstract::Accumulating evidence has indicated that microRNAs (miRNAs) act as critical epigenetic regulators in tumor carcinogenesis. Here, we report that miR-199a-3p was significantly upregulated in gastric cancer (GC) cell lines and tissues. Functional studies demonstrated that miR-199a-3p dramatically increased cell prolifera...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2014.09.047
更新日期:2014-11-28 00:00:00
abstract::We have isolated a full-length cDNA (HPAsn.6) for human placenta glycosylasparaginase using a 221-bp PCR amplified fragment containing rat liver asparaginase gene sequences. The deduced amino acid sequence from the human clone showed sequence identity to both the alpha and beta subunits of the rat enzyme. The human en...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(90)81211-6
更新日期:1990-09-03 00:00:00
abstract::We tested the effect of melatonin on membrane fluidity in microsomes of a rat liver model in which lipid peroxidation was induced by the addition of FeCl3, ADP and NADPH. Membrane fluidity was monitored using fluorescence spectroscopy and lipid peroxidation was estimated by quantifying malonaldehyde (MDA)+4-hydroxyalk...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(97)00447-x
更新日期:1997-05-26 00:00:00
abstract::Treatment of cultured carrot cells with dibutyryl cAMP or forskolin resulted in the appreciable decrease in extracellular K+ concentration. This decrease was found to be transient and the concentration of the ion in the culture medium restored to the original level within few minutes. The cAMP-induced decrease in K+ l...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(97)00403-1
更新日期:1997-05-12 00:00:00
abstract::Apart from the 8 kDa subunit, which is of chloroplast origin, most of the small polypeptides of the PS I reaction center from higher plants are encoded in nuclear genes. We describe here the first nucleotide sequence of a nuclear component of this photosystem, the precursor of the 20 kDa protein. The deduced sequence ...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(88)80752-x
更新日期:1988-05-23 00:00:00
abstract::Protein C (PC) is a vitamin K-dependent zymogen that inactivates factors Va and VIIIa after its activation by thrombin complexed to thrombomodulin. We characterized a monoclonal antibody (mAb) against PC, whose only influence on PC functions was to inhibit PC activation by the thrombin-thrombomodulin complex. It recog...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(95)00552-k
更新日期:1995-06-26 00:00:00
abstract::The expression of myf-5, a key component of myogenic regulatory genes, declines progressively in mature somitic cells during vertebrate myogenesis. Little is known about how this down-regulation takes place. Here we provide evidence that an interferon regulatory factor binding element (IRF element) within the Xenopus ...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(01)02688-6
更新日期:2001-09-07 00:00:00
abstract::The dependence of protein splicing on conserved residues of the Cne PRP8 intein was assessed by alanine scanning mutagenesis in a foreign protein context. Corroboration was obtained for the involvement of residues at the splice junctions and of the conserved threonine and histidine of motif B. Five additional residues...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2007.05.060
更新日期:2007-06-26 00:00:00
abstract::The phylogenetic distribution of divergently related protein families into the three domains of life (archaea, bacteria and eukaryotes) can signify the presence or absence of entire cellular processes in these domains and their ancestors. We can thus study the emergence of the major transitions during cellular evoluti...
journal_title:FEBS letters
pub_type: 杂志文章,评审
doi:10.1016/0014-5793(96)00631-x
更新日期:1996-07-22 00:00:00
abstract::Dun1p and Rad53p of the budding yeast Saccharomyces cerevisiae are members of a conserved family of cell cycle checkpoint protein kinases that contain forkhead-associated (FHA) domains. Here, we demonstrate that these FHA domains contain 130-140 residues, and are thus considerably larger than previously predicted by s...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(00)01392-2
更新日期:2000-04-14 00:00:00
abstract::Gastrin (G17) and N-carboxymethylgastrin (G17-Gly) have been shown to stimulate the growth of colon cancer cells both in vivo and in vitro. The identity of the receptor mediating these effects is controversial. A recent study demonstrated the presence of a low affinity binding site for G17 and G17-Gly on the DLD-1 hum...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(03)01408-x
更新日期:2004-01-02 00:00:00
abstract::Insertion and deletion analyses of a protein have been less common than point mutation analyses, partly due to the lack in effective methods. This is the case with the green fluorescent protein (GFP), which is so widely applied in molecular biology and other fields. In this paper we first introduce a systematic approa...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(03)01308-5
更新日期:2003-12-18 00:00:00
abstract::The Escherichia coli twin-arginine translocation (Tat) system serves to export fully folded protein substrates across the bacterial cytoplasmic membrane. Respiratory [NiFe] hydrogenases are synthesised as precursors with twin-arginine signal peptides and transported as large, cofactor-containing, multi-subunit complex...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(03)00802-0
更新日期:2003-08-14 00:00:00
abstract::During the course of large scale purification of the D1 dopamine receptor from rat brain, a protein of approximately 87,000 daltons (p87) was observed to copurify with the D1 receptor through four chromatographic steps. To characterize the nature of this protein, bovine and rat cDNA clones were isolated and sequenced....
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(92)80917-6
更新日期:1992-11-09 00:00:00
abstract::The nucleotide sequence of the Pseudomonas saccharophila gene encoding maltotetraohydrolase (G4-forming amylase) has been determined. The coding region for the G4-forming amylase precursor contained 1653 nucleotides. The deduced precursor protein included an N-terminal 21-residue putative signal peptide; the deduced m...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(89)81056-7
更新日期:1989-09-11 00:00:00
abstract::Placental alkaline phosphatase is an inducible enzyme, expressed in HeLaS3 cells, which has been shown to possess protein phosphotyrosine phosphatase activity. Since phosphotyrosine levels are known to increase in actively dividing cells we sought an inverse correlation between PLAP activity and growth rate in HeLaS3 ...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(93)80229-n
更新日期:1993-08-30 00:00:00
abstract::The NAD+-requiring enzymes of glycoside hydrolase family 4 (GHF4) contain a region with a conserved Gly-XXX-Gly-Ser (GXGS) motif near their N-termini that is reminiscent of the fingerprint region of the Rossmann fold, a conserved structural motif of classical nicotinamide nucleotide-binding proteins. The function of t...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(02)02641-8
更新日期:2002-04-24 00:00:00
abstract::Exposing a macrophage-like murine cell line to copper-oxidised low-density lipoprotein led to DNA fragmentation which was inhibited by the putative Ca2+/Mg2+ endonuclease inhibitor, zinc sulphate. DNA fragmentation preceded loss of membrane impermeability. These results suggest that apoptosis may be a mechanism of mac...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(93)80635-8
更新日期:1993-10-18 00:00:00
abstract::Farnesoic acid is a signaling molecule that inhibits the transition from budding yeast to filament formation in Candida albicans, but the molecular mechanism regulated by this substance is unknown. In this study, we analyzed the function of CaPHO81, which is induced by farnesoic acid. The pho81Δ mutant cells existed e...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2010.10.026
更新日期:2010-11-19 00:00:00
abstract::The role of protein kinase C (PKC) and p42(MAP kinase) signaling in the regulation of proliferation and apoptosis was investigated in freshly isolated and primary cultured rat hepatocytes. Acute treatment of freshly isolated hepatocytes with phenylephrine and EGF caused rapid phasic activations of p42(MAP kinase) and ...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(97)00705-9
更新日期:1997-07-21 00:00:00
abstract::MM-1 has been reported to repress the E-box-dependent transcription activity of c-Myc by recruiting histone deacetylase 1 complex via TIF1beta/KAP1. In this study, to identify target genes for c-Myc-MM-1-TIF1beta, we established rat-1 cells harboring the dominant-negative form of TIF1beta to abrogate the pathway from ...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2004.07.034
更新日期:2004-08-13 00:00:00
abstract::Four 12.2-12.6 kDa small heat-shock proteins (sHSPs) of Caenorhabditis elegans are the smallest known members of the sHSP family. They essentially comprise the characteristic C-terminal 'alpha-crystallin domain' of the sHSPs, having a very short N-terminal region, and lacking a C-terminal tail. Recombinant Hsp12.2 and...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/s0014-5793(98)00917-x
更新日期:1998-08-21 00:00:00
abstract::We have previously identified an ESE in NF1 exon 37 whose disruption by the pathological mutation c.6792C>G caused aberrant splicing. We now investigate the RNA-protein complexes affected by the c.6792C>G mutation observing that this concurrently decreases the affinity for the positive splicing factor YB-1 and increas...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2008.05.018
更新日期:2008-06-25 00:00:00
abstract::Chicken muscular dystrophy with abnormal muscle (AM) has been studied for more than 50 years, but the gene responsible for it remains unclear. Our previous studies narrowed down the AM candidate region to approximately 1Mbp of chicken chromosome 2q containing seven genes. In this study, we performed sequence compariso...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2008.05.013
更新日期:2008-06-25 00:00:00
abstract::It is known that narcotic antagonists interact with many cholinergic pathways but less in known about specific mechanisms. Using neonatal rat myoballs to study single channel behaviour of the acetylcholinegated nicotinic receptor, it was found that micromolar concentrations of naltrexone had no effect on channel condu...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(85)81145-5
更新日期:1985-03-11 00:00:00
abstract::Hepatitis C virus (HCV) replication and pathogenesis involve both virus-encoded proteins and cellular factors. In our study, we showed that NS5B, the HCV RNA-dependent RNA polymerase, interacted with M2 type pyruvate kinase (M2PK) but not L type pyruvate kinase. We confirmed the interaction by GST pull down, coimmunop...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2008.05.033
更新日期:2008-06-25 00:00:00
abstract::The diffusion of fluorescein isothiocyanate-labelled dextran molecules in suspensions of centrifugally, tightly packed, erythrocyte ghosts was measured by fluorescence recovery after photobleaching. In comparison with diffusion in aqueous solution, the diffusion coefficients for probe molecules of varying size were ab...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(88)81000-7
更新日期:1988-12-19 00:00:00
abstract::The substrate peptide bond cleaved by cathepsins B and L is determined not by the amino acid contributing the carboxyl group to this bond as in the case of serine proteases but rather by the presence of a neighboring amino acid with a large hydrophobic side chain. From a study of the inhibitory potency in a series, Cb...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/0014-5793(88)80600-8
更新日期:1988-02-08 00:00:00
abstract::Adrenomedullin (ADM) functions as a survival factor against hypoxic cell death. However, molecular mechanisms underlying the cell survival pathway remain largely unknown. In this report, we showed that ADM suppressed reactive oxygen species (ROS) increase by inhibiting reduction of glutathione (GSH) level in hypoxia/r...
journal_title:FEBS letters
pub_type: 杂志文章
doi:10.1016/j.febslet.2009.11.063
更新日期:2010-01-04 00:00:00