Cold adaptation parameters derived from cDNA sequencing and molecular modelling of elastase from Antarctic fish Notothenia neglecta

Aittaleb, Mohamed; Hubner, Romeo J.; Lamotte‐Brasseur, Josette; Gerday, Charles

doi:10.1093/protein/10.5.475

Cited by 33 publications

(19 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This is the case with the Trp 336 displacement in native AgaB that widens the catalytic cleft. This larger opening of the active site was demonstrated, for example, in the structures of Antarctic fish elastase (39) and cold-active citrate synthase (40). The gain in accessibility can help to reduce the energy required for substrate accommodation and/or reaction product release in the case of macromolecular substrates.…”

Section: Site-directed Mutational and Kinetic Studies-we Further Invementioning

confidence: 92%

The Molecular Mechanism of Thermostable α-Galactosidases AgaA and AgaB Explained by X-ray Crystallography and Mutational Studies

Merceron

Foucault

Haser

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The thermostable ␣-galactosidases AgaA and AgaB are of industrial interest and have different catalytic properties despite having a high sequence identity. Results: The crystal structures helped to explain the specificities of these two isoenzymes. Conclusion: The A355E substitution modulates the induced fit mechanism. Significance: Transglycosylation is increased by a single substitution that is located away from the active site.

show abstract

Section: Site-directed Mutational and Kinetic Studies-we Further Invementioning

confidence: 92%

The Molecular Mechanism of Thermostable α-Galactosidases AgaA and AgaB Explained by X-ray Crystallography and Mutational Studies

Merceron

Foucault

Haser

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Pairwise sequence alignment was performed on the Genestream server at http ://genome.eerie.fr/ bin/align-guess.cgi. The sequences used for alignment were: Atlantic cod elastase B (present report and TREMBL Q92077), Atlantic cod chymotrypsin variant A (Swiss-Prot P47796), variant B (Swiss-Prot P80646), Atlantic cod elastase A (TREMBL Q91039), Antarctic fish Notothenia neglecta elastase [20], salmon elastase (PDB 1ELT), porcine elastase 1 (Swiss-Prot P00772), human elastase 2A (Swiss-Prot P08217), and rat elastase 2 (Swiss-Prot P00774).…”

Section: Methodsmentioning

confidence: 99%

“…Atlantic cod elastase I did not hydrolyse Suc-AAPF-NH-Np but was active towards Suc-AAA-NH-Np and elastin-orcein [17]. Most recently, the primary structure of an elastase from the Antarctic fish N. neglecta was elucidated [20]. The amino acid sequence identity with mammalian elastases ranged over 53Ϫ64%, but reached 79% in comparison with cod chymotrypsin B.…”

Section: Kinetic Properties Of Elastase C and The Effect Of Inhibitorsmentioning

confidence: 99%

The third serine proteinase with chymotrypsin specificity isolated from Atlantic cod (Gadus morhua) is a type‐II elastase

Ásgeirsson

Leth-Larsen²,

Thórólfsson

et al. 1998

European Journal of Biochemistry

View full text Add to dashboard Cite

Preparations of chymotrypsin from Atlantic cod are heterogeneous and previously gave rise to two active peaks when subjected to pH-gradient chromatography. Extension of the pH-gradient resolved a third protein peak with benzoyltyrosine ethylester hydrolytic activity. The first two peaks have been characterized as chymotrypsin variants and designated A and B, whereas the identity of the third peak was not clear. Analysis of this protein by Edman sequencing and mass spectrometry has now confirmed a high degree of identity with the predicted protein sequence from a recently described cDNA clone. That sequence was named elastase B by sequence comparison. As the present elastase deviates in 16 positions from that of elastase B, we have named it elastase C. The elastase C was active in hydrolysing typical substrates used by chymotrypsin, namely benzoyl-L-tyrosine ethylester and succinyl-Ala-Ala-ProPhe-p-nitroanilide, but inactive against the typical elastase substrates succinyl-Ala-Ala-Ala-p-nitroanilide and orcein-elastin. Comparison of the kinetic properties of the cod elastase C with bovine chymotrypsin and cod chymotrypsin variants A and B, using succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, showed a lower catalytic efficiency of elastase C. The effects of several inhibitors on cod elastase C were identical to effects on chymotrypsins variants A and B, but dissimilar when compared with porcine pancreatic elastase. On the basis of the specificity and amino acid sequence, we conclude that the enzyme under study is most correctly classified as a type-II elastase.Keywords : elastase; serine protease ; Atlantic cod ; amino acid sequence; cold adaptation.Variants of elastases have been identified in different animals and in different animal tissues such as the pancreas, leucocytes, platelets, and aorta [1]. Elastase was initially characterized as a mammalian pancreatic serine protease with the ability to dissolve elastin [2], but elastase can digest a wide variety of other protein substrates in addition to native elastin [3]. Some elastases in the serine proteinases family turn out to be specific for non-bulky amino acid residues (e.g. porcine pancreatic elastase 1, human leucocyte elastase) while others have a chymotrypsinlike specificity (e.g. porcine pancreatic elastase 2, human pancreatic elastase 2). Pancreatic elastase type I was originally isolated from hog pancreas and is strongly cationic with an isoelectric point near pH 9.5 [2]. Another anionic enzyme, called elastase type II, was also obtained from the same tissue and found to have a characteristic chymotrypsin activity [4,5]. Human pancreatic tissue and juice also contain two enzymes, one cationic and the other anionic, which can hydrolyse a specific substrate for porcine pancreatic elastase and undyed elastin [6,7]. The cationic enzyme is the major elastase in human pancreas and has similar properties to porcine pancreatic elastase 1, whereas elastase 2 preferentially hydrolysed peptides at sites of medium to large hydrophobic side chains [8]. Human elastase 1 ...

show abstract

“…In addition to the above mentioned crystal structures [50,51,52] models of psychrophilic enzymes such as bacterial subtilisin [59], h-amylase [45], triosephosphate isomerase [60], lipase [61], i-lactamase [48] or fish trypsin [62] and elastase [63] reveal that only subtle modifications of their conformation can be related to the low stability (see also [64,65]). Electrostatic weak interactions and the hydrophobic effect make a lower contribution to the global energy of stabilization of the native state whereas destabilizing forces mainly favour the conformational entropy of the unfolded state.…”

Section: Structural Factors Affecting the Stability Of Psychrophilic mentioning

confidence: 99%

“…For instance, the low stability of the psychrophilic subtilisin was tentatively correlated with the occurrence of several long surface loops which improve interactions with the solvent and lower the compactness of the molecular surface [59]. In the case of elastase from the Antarctic fish N. neglecta, a small deletion make the entrance of the catalytic cavity markedly more accessible, possibly allowing easier substrate accommodation [63]. Two small insertions in loops involved in dimerization of Moraxella triosephosphate isomerase were also expected to favour dimer destabilization [60].…”

Section: Structural Factors Affecting the Stability Of Psychrophilic mentioning

confidence: 99%

Psychrophilic enzymes: molecular basis of cold adaptation

Feller

Gerday

1997

CMLS, Cell. mol. life sci.

Self Cite

370

233

View full text Add to dashboard Cite

Abstract. Psychrophilic organisms have successfully provides enhanced abilities to undergo conformational colonized polar and alpine regions and are able to grow changes during catalysis. Thermal instability of coldefficiently at sub-zero temperatures. At the enzymatic adapted enzymes is therefore regarded as a consequence level, such organisms have to cope with the reduction of of their conformational flexibility. A survey of the psychrophilic enzymes studied so far reveals only minor chemical reaction rates induced by low temperatures in alterations of the primary structure when compared to order to maintain adequate metabolic fluxes. Thermal compensation in cold-adapted enzymes is reached mesophilic or thermophilic homologues. However, all known structural factors and weak interactions inthrough improved turnover number and catalytic effivolved in protein stability are either reduced in number ciency. This optimization of the catalytic parameters can originate from a highly flexible structure which or modified in order to increase their flexibility.Key words. Psychrophiles; extremophiles; cold adaptation; microbial proteins; protein stability; weak interactions; Antarctic.Psychrophiles live at the lowest temperatures which allow the development of living organisms. These extremophilic organisms, either prokaryotic or eukaryotic, represent a major class of the living world if we consider the vast extent of permanently cold environments on Earth, such as polar and alpine regions or deep-sea waters. The successful colonization of such severe ecological niches by psychrophiles, their diversity and abundance are now well recognized. The lower temperature limit of life is commonly defined as the freezing point of cellular water. Although apparently simple by definition, this limit can drop significantly below 0°C, the freezing point of pure water. For instance, the freezing point of a typical marine teleost ranges between −0.5 and −0.9°C. Sodium chloride is responsible for about 85% of the freezing point depression, and the remaining 15% is due to other small solutes. Synthesis of antifreeze glycoproteins and peptides can further depress the freezing point of body fluids or cellular water. These proteins lower the freezing point by a noncolligative mechanism without altering the melting point significantly, and are therefore also referred to as thermal hysteresis proteins [1,2]. They were first discovered in the blood sera of Antarctic fish living perennially at about −1.9°C, but have been now reported in many organisms including plants, insects, fungi and bacteria [3][4][5]. Levels of thermal hysteresis activity generally range from 0.1 to 0.3°C in bacteria, from 0.2 to 0.5°C in plants, from 0.7 to 1.5°C in fish and from 3 to 6°C in insects. Besides the synthesis of cryoprotectants which lead to freeze avoidance, several organisms have developed mechanisms of freeze tolerance, involving drastic metabolic modifica-* Corresponding author.

show abstract

Cold adaptation parameters derived from cDNA sequencing and molecular modelling of elastase from Antarctic fish Notothenia neglecta

Cited by 33 publications

References 0 publications

The Molecular Mechanism of Thermostable α-Galactosidases AgaA and AgaB Explained by X-ray Crystallography and Mutational Studies

The Molecular Mechanism of Thermostable α-Galactosidases AgaA and AgaB Explained by X-ray Crystallography and Mutational Studies

The third serine proteinase with chymotrypsin specificity isolated from Atlantic cod (Gadus morhua) is a type‐II elastase

Psychrophilic enzymes: molecular basis of cold adaptation

Contact Info

Product

Resources

About