Cystatin. Amino acid sequence and possible secondary structure

Schwabe, Christian; Xuereb-Anastasi, Angela; Crow, H. J.; McDonald, J. Ken; Barrett, Alan J.

doi:10.1042/bj2170813

Cited by 83 publications

(50 citation statements)

References 13 publications

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“…The most highly conserved residue Gly 9 (chicken cystatin numbering) found in all known sequences of inhibitory cystatins [14], bovine colostrum cystatin (e) [15], and chicken cystatin (f) [16,17]. Residues are numbered according to chicken cystatin.…”

Section: Resultsmentioning

confidence: 99%

Primary structure of a new cysteine proteinase inhibitor from pig leucocytes

et al. 1989

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The primary structure of a pig leucocyte cysteine proteinase inhibitor, also called cathelin, was determined. The sequence was obtained from analyses of peptides isolated from the chymotryptic, endoproteinase Lys-C and protease V8 digests, and by analysis of the peptides derived from the hydrolysis of the aspartyl-prolyl bond of the carboxymethylated inhibitor. The inhibitor consists of 96 residues. The N-terminal residue of the inhibitor is pyrrolidonecarboxylic acid. The amino acid sequence. of cathelin suggests the appearance of a new family of cysteine proteinase inhibitors.

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Section: Resultsmentioning

confidence: 99%

Primary structure of a new cysteine proteinase inhibitor from pig leucocytes

et al. 1989

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“…guanidinobutane; FPLC, fast protein liquid chromatography [4,6]. Also shorter forms starting with Gly-9 (Glyform) and Ala-10 (Ala-form) have been found [4,7].…”

Section: Abbreviations: E-64 1-(trans-epoxysuccinyl-l-leucylamino)-4mentioning

confidence: 99%

Mechanism of inhibition of papain by chicken egg white cystatin

et al. 1989

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N-terminally truncated forms of chicken egg white cystatin and its cyanogen bromide fragments were isolated and assayed for inhibition of papain. Truncated forms beginning with Gly-9 and Ala-10 had a 5000-fold lower affinity for papain than the two isoelectric forms (pi=6.5 and 5.6) of the full-length inhibitor (K~--6 pM and 7 pM) or a truncated form beginning with Leu-7 (/~ = 6 pM), indicating the outstanding importance of one or two residues preceding conserved Gly-9 for binding. A weak inhibition of papain (K~ = 900 nM) was exhibited by the intermediate cyanogen bromide fragment (residues 30-89) containing the chicken cystatin QLVSG variation of the QWAG segment which is conserved in almost all members of the cystatin superfamily. The obtained affinity data provide independent evidence for the validity of the proposed docking model of a chicken cystatin-papain complex [( ) EMBO J. 7, 2593[( -2599.

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“…3 b. According to the known chicken cystatin sequence [3,5], we expected identification problems with the short tryptic peptides Tp2, Tp9 and Tp16. Sequence analysis of Tp2 was performed by Nterminal sequencing, whereas that of Tp9 was determined by sequencing a fusion peptide of Tp9 and TplO.…”

Section: Primary Structurementioning

confidence: 99%

“…Affinity chromatography was performed essentially as described [4,5,211. Material used for NMR studies (about 5 -10 mg) was desalted by gel filtration on PD-10 columns (Pharmacia) according to the instructions of the manufacturer.…”

Section: Cultivation Of E Coli and Isolation Of Recombinant Cystatinmentioning

confidence: 99%

Purification and characterization of a chicken egg white cystatin variant expressed in an Escherichia coli pIN‐III‐ompA system

Auerswald¹,

Genenger²,

Mentele³

et al. 1991

European Journal of Biochemistry

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A synthetic gene coding for a chicken egg white cystatin variant was cloned and expressed using the pIN‐III‐ompA Escherichia coli expression system. After osmotic shock of the E. coli cells, the cysteine proteinase inhibitor was isolated from periplasm and purified by S‐carboxymethylpapain affinity chromatography. The resulting inhibitory material was characterized by SDS/PAGE, reversed‐phase HPLC, peptide mapping and amino acid sequencing. The recombinant variant chicken AEF‐[S1 → M, M29 → I, M89 → L]cystatin shows strong inhibitory activity and displays Ki values in the complex with papain, actinidin and cathepsin B similar to those found for natural chicken cystatin. The purified variant showed a native‐chicken‐cystatin‐like conformational state, as determined by NMR spectroscopy, if the NMR data of 15N‐labelled recombinant inhibitor were compared with those of the natural inhibitor.

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Cystatin. Amino acid sequence and possible secondary structure

Cited by 83 publications

References 13 publications

Primary structure of a new cysteine proteinase inhibitor from pig leucocytes

Primary structure of a new cysteine proteinase inhibitor from pig leucocytes

Mechanism of inhibition of papain by chicken egg white cystatin

Purification and characterization of a chicken egg white cystatin variant expressed in an Escherichia coli pIN‐III‐ompA system

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