Nomenclature and classification of the proteins homologous with the cysteine-proteinase inhibitor chicken cystatin

Barrett, A J; Fritz, Hans; Grubb, Anders; Isemura, Satoko; Järvinen, Mikko; Katunuma, Nobuhiko; Machleidt, Werner; Müller‐Esterl, Werner; Sasaki, Makoto; Türk, Vito

doi:10.1042/bj2360312

Cited by 298 publications

(129 citation statements)

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“…Release of the product was continuously recorded and typical biphasic progress curves were Eight members of the stefin family [2] of mammalian cysobserved. Measurements were analysed by fitting the product teine proteinase inhibitors have been characterized on the release data to the integrated rate equation [28].…”

Section: Discussionmentioning

confidence: 99%

Differences in specificity for the interactions of stefins A, B and D with cysteine proteinases

et al. 1996

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Four different stefin-type cysteine proteinase inhibitin [15,16] and of a complex of stefin B with carboxymethyltors have been isolated from porcine thymus and skin. Amino papain [17]. The reactive site of the inhibitor comprises of two acid sequence determination revealed the presence of stefin A and hairpin loops and the N-terminal part of the inhibitor forms a stefin B type inhibitors and two new inhibitors, designated as hydrophobic wedge-shaped edge, which is highly complemenporcine stefin D1 and stefin D2. Stefin D1 was identified as tary to the active site of the papain. In this paper we present the purification and characterizacathepsin S (Ki =0.009-0.161 nM). Stefins A and B also bind tightly and rapidly to cathepsin H (Ki = 0.027 and 0.069 nM, tion from porcine tissue of stefins A and B and two new respectively), while stefins D1 and D2 have been shown to be very stefin-type inhibitors, designated stefins D1 and D2. The cornpoor inhibitors of cathepsin H (Ki = 102-150 nM). The decreased plete amino acid sequences of stefins A, B and D1 were deaffinity of these inhibitors toward cathepsin B (Ki = 2-1700 nM) termined and compared with other known stefin-type sewas shown to be mainly due to the low second order association quences. In addition, their kinetics of interaction with rate constants. The presence of a highly negatively charged Npapain-like cysteine proteinases were determined. terminus on stefin D1 constitutes a likely structural determinant of inhibitor specificity. Materials and methods

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Section: Discussionmentioning

confidence: 99%

Differences in specificity for the interactions of stefins A, B and D with cysteine proteinases

et al. 1996

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“…In their sequence [l-31, there are more than 50% identical amino acid residues. Human stefins A and B are representatives of the stefin family of the cystatin superfamily of the cysteine-proteinases inhibitors [4,5]. The three-dimensional (3D) structure of a closely related cysteine-proteinase inhibitor, the chicken egg-white cystatin, has been determined by Bode and coworkers [6].…”

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confidence: 99%

Calorimetric measurements of thermal denaturation of stefins A and B

Žerovnik

Lohner

Jerala

et al. 1992

European Journal of Biochemistry

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Thermal denaturation of two homologous proteins, low-M, cysteine-proteinase inhibitors stefins A and B, has been investigated by microcalorimetry. Calorimetric enthalpies, as well as the temperatures at maximum heat capacity, were determined as a function of pH for each protein. Transitions were found reversible at all pH values examined (5.0, 6.5, 8.1) for the thermally more stable stefin A, in contrast to stefin B. Stefin B shows a sharp irreversible transition around 65°C at pH 6.5 and 8.1, probably due to unfolding of a dimeric state followed by oligomerisation. At pH 5.0, both proteins exhibit a reversible transition with temperatures of half-denaturation at 50.2 "C and 90.8 "C for stefins B and A, respectively. The calorimetric enthalpies, which equal the van? Hoff enthalpies to within lo%, are 293 kJ/mol and 490 kJ/mol for stefins B and A, respectively.Using the predictive method of Ooi and Oobatake (1991) [Proc. Natl Acad. Sci. USA 88, 28591 the thermodynamic functions of unfolding were calculated for stefin B, whose three-dimensional structure has been determined. The calculated enthalpy, heat-capacity change on unfolding and the temperature of half denaturation compare well to the microcalorimetric data.Human stefins A and B are small globular proteins of M , 11 000, without disulphide bonds. In their sequence [l-31, there are more than 50% identical amino acid residues. Human stefins A and B are representatives of the stefin family of the cystatin superfamily of the cysteine-proteinases inhibitors [4,5]. The three-dimensional (3D) structure of a closely related cysteine-proteinase inhibitor, the chicken egg-white cystatin, has been determined by Bode and coworkers [6]. The same group determined the 3D structure of the complex between papain and stefin B [7].Spectroscopic study of Gdn/HCl, heat and pH denaturation of the recombinant human stefin B [8] has shown that this protein is much less stable than stefin A. Equilibrium intermediates of molten globular type could be detected in the stefin-B denaturation [8], whereas stefin A seems to denature in an all-or-nothing manner [9].Difference scanning calorimetry yields thermodynamic data on the stability of proteins [lo-121. The temperature dependence of the enthalpy and the entropy of unfolding can be calculated if the change in heat capacity on unfolding is determined. It is a long-standing goal to be able to predict the thermodynamic functions of unfolding as this could contribute towards a rational design of new proteins. A semiempirical method of Ooi and Oobatake [13-151 permits such a calculation from the difference in the accessible surface areas between denatured and native states of a protein, respectively.Correspondence to E. ierovnik, Department of Biochemistry, Fax: f 3 8 61 273 594. Abbreviation. 3D, three-dimensional.Joief Stefkn Institute, Jamova 39, Ljubljana, Slovenia 61 000Our aim was to study the difference in thermal denaturation of the two homologous proteins, of which stefin A shows quite a respective thermal stability and stefin B ...

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“…BI has no similarity to the cystatin superfamily inhibitors in the primary structure (1) and has no Gln-X-Val-X-Gly sequence that is essential for activity against papain and other cysteine proteinases (7). For example, the structure of BI-VI N shows no homology with that of rice cystatin that consists of an ␣-helix and a five-stranded antiparallel ␤-sheet (8).…”

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confidence: 99%

“…Most cysteine proteinase inhibitors obtained from plants so far have been identified as cystatins including inhibitors from rice, corn, soybean, and tomato (1). Their properties have been extensively studied, including their nucleotide and amino acid sequences and three-dimensional structures.…”

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confidence: 99%

“…Their properties have been extensively studied, including their nucleotide and amino acid sequences and three-dimensional structures. Reddy et al (2) have isolated a novel and microheterogeneous inhibitor toward bromelain (BI) 1 from pineapple (Ananas comosus) stem (2). Each isoform of BI consists of a light chain (10 or 11 residues) and a heavy chain (40 or 41 residues) that are cross-linked with disulfide bonds (2)(3)(4).…”

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Characterization of Genomic Sequence Coding for Bromelain Inhibitors in Pineapple and Expression of Its Recombinant Isoform

Sawano¹,

Muramatsu²,

Hatano³

et al. 2002

Journal of Biological Chemistry

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Bromelain inhibitor (BI) is a cysteine proteinase inhibitor isolated from pineapple stem (Reddy, M. N., Keim, P. S., Heinrikson, R. L., and Ké zdy, F. J. (1975) J. Biol. Chem. 250, 1741-1750). It consists of eight isoinhibitors, and each isoinhibitor has a two-chain structure. In this study, the genomic DNA has been cloned and found to encode a precursor protein with 246 amino acids (M r ‫؍‬ ϳ27,500) containing three isoinhibitor domains (BI-III, -VI, and -VII) that are 93% identical to one another in amino acid sequences. The gene structure indicated that these isoinhibitors are produced by removal of the N-terminal pre-peptide (19 residues), 3 interchain peptides (each 5 residues), 2 interdomain peptides (each 19 residues), and the C-terminal pro-peptide (18 residues). Moreover, all the amino acid sequences of bromelain isoinhibitors could be explained by removal of one or two amino acids from BI-III, -VI, and -VII with exopeptidases. A recombinant single-chain BI-VI with and without the interchain peptide showed the same and no bromelain inhibitory activity as compared with the native BI-VI, respectively. These results indicate that the interchain peptide plays an important role of the folding process of the mature isoinhibitors.Most cysteine proteinase inhibitors obtained from plants so far have been identified as cystatins including inhibitors from rice, corn, soybean, and tomato (1). Their properties have been extensively studied, including their nucleotide and amino acid sequences and three-dimensional structures. Reddy et al. (2) have isolated a novel and microheterogeneous inhibitor toward bromelain (BI) 1 from pineapple (Ananas comosus) stem (2). Each isoform of BI consists of a light chain (10 or 11 residues) and a heavy chain (40 or 41 residues) that are cross-linked with disulfide bonds (2-4). The three-dimensional solution structure of the sixth isoinhibitor (BI-VI N ) has two distinct domains named A and B domains, each of which is formed by a threestranded antiparallel ␤-sheet (5, 6). The B domain consists of the light chain and part of the heavy chain (Glu 1H -Asp 9H and Leu 31H -Lys 41H ) 2 and was indicated to involve the major bromelain inhibitory site, possessing a more flexible structure that seems to allow the inhibitor to fit well into the catalytic site region of a target proteinase (4, 6). On the other hand, the structure of the A domain (Thr 10H -Ile 29H ) was well converged and, thus, thought to contribute greatly to the conformational stability of the B domain (6).BI has no similarity to the cystatin superfamily inhibitors in the primary structure (1) and has no Gln-X-Val-X-Gly sequence that is essential for activity against papain and other cysteine proteinases (7). For example, the structure of BI-VI N shows no homology with that of rice cystatin that consists of an ␣-helix and a five-stranded antiparallel ␤-sheet (8). Interestingly, BI-VI N shared the same folding and disulfide bond connectivities with Bowman-Birk serine proteinase inhibitor (BBI) from soybean (6), a 71-residue in...

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Nomenclature and classification of the proteins homologous with the cysteine-proteinase inhibitor chicken cystatin

Cited by 298 publications

References 1 publication

Differences in specificity for the interactions of stefins A, B and D with cysteine proteinases

Differences in specificity for the interactions of stefins A, B and D with cysteine proteinases

Calorimetric measurements of thermal denaturation of stefins A and B

Characterization of Genomic Sequence Coding for Bromelain Inhibitors in Pineapple and Expression of Its Recombinant Isoform

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