scholarly journals α2 -Macroglobulin bait region integrity

FEBS Letters ◽  
1993 ◽  
Vol 325 (3) ◽  
pp. 267-270 ◽  
Author(s):  
Peter G.W. Gettins ◽  
Joseph M. Beechem ◽  
Brenda C. Crews
Keyword(s):  
Bait Region ◽  
Α2 Macroglobulin

scholarly journals Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin

2021 ◽  
pp. 100879
Author(s):  
Seandean Lykke Harwood ◽  
Nadia Sukusu Nielsen ◽  
Khang Diep ◽  
Kathrine Tejlgård Jensen ◽  
Peter Kresten Nielsen ◽  
...  
Keyword(s):  
Protease Inhibitors ◽  
Bait Region ◽  
Α2 Macroglobulin

scholarly journals Structural and functional insights into Escherichia coli α2-macroglobulin endopeptidase snap-trap inhibition

2015 ◽  
Vol 112 (27) ◽  
pp. 8290-8295 ◽  
Author(s):  
Irene Garcia-Ferrer ◽  
Pedro Arêde ◽  
Josué Gómez-Blanco ◽  
Daniel Luque ◽  
Stephane Duquerroy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Envelope ◽  
Large Cell ◽  
Structural Rearrangement ◽  
Gram Negative Bacteria ◽  
Human Gut ◽  
Bait Region ◽  
Α2 Macroglobulin ◽  
Thioester Bond ◽  
Peptidase Inhibitor

The survival of commensal bacteria requires them to evade host peptidases. Gram-negative bacteria from the human gut microbiome encode a relative of the human endopeptidase inhibitor, α2-macroglobulin (α2M). Escherichia coli α2M (ECAM) is a ∼180-kDa multidomain membrane-anchored pan-peptidase inhibitor, which is cleaved by host endopeptidases in an accessible bait region. Structural studies by electron microscopy and crystallography reveal that this cleavage causes major structural rearrangement of more than half the 13-domain structure from a native to a compact induced form. It also exposes a reactive thioester bond, which covalently traps the peptidase. Subsequently, peptidase-laden ECAM is shed from the membrane and may dimerize. Trapped peptidases are still active except against very large substrates, so inhibition potentially prevents damage of large cell envelope components, but not host digestion. Mechanistically, these results document a novel monomeric “snap trap.”

scholarly journals Primary structure of the ‘bait’ region for proteinases in α2 -macroglobulin

FEBS Letters ◽  
1981 ◽  
Vol 127 (2) ◽  
pp. 167-173 ◽  
Author(s):  
Lars Sottrup-Jensen ◽  
Peter B. Lønblad ◽  
Terrence M. Stepanik ◽  
Torben E. Petersen ◽  
Staffan Magnusson ◽  
...  
Keyword(s):  
Primary Structure ◽  
Bait Region ◽  
Α2 Macroglobulin

scholarly journals Pig kidney legumain: an asparaginyl endopeptidase with restricted specificity

1999 ◽  
Vol 339 (3) ◽  
pp. 743-749 ◽  
Author(s):  
Pam M. DANDO ◽  
Mara FORTUNATO ◽  
Lorraine SMITH ◽  
C. Graham KNIGHT ◽  
John E. MCKENDRICK ◽  
...  
Keyword(s):  
Mhc Class Ii ◽  
Tetanus Toxin ◽  
Tertiary Butyl ◽  
Asparaginyl Endopeptidase ◽  
Pig Kidney ◽  
Bait Region ◽  
Substrate Analogues ◽  
Α2 Macroglobulin ◽  
Hydrolysis Of ◽  
Asparagine Residue

Legumain was recently discovered as a lysosomal endopeptidase in mammals [Chen, Dando, Rawlings, Brown, Young, Stevens, Hewitt, Watts and Barrett (1997) J. Biol. Chem. 272, 8090-8098], having been known previously only from plants and invertebrates. It has been shown to play a key role in processing of the C fragment of tetanus toxin for presentation by the MHC class-II system [Manoury, Hewitt, Morrice, Dando, Barrett and Watts (1998) Nature (London) 396, 695-699]. We examine here the specificity of the enzyme from pig kidney by use of protein, oligopeptide and synthetic arylamide substrates, all determinations being made at pH 5.8. In proteins, only about one in ten of the asparaginyl bonds were hydrolysed, and these were mostly predicted to be located at turns on the protein surface. Bonds that were not cleaved in tetanus toxin were cleaved when presented in oligopeptides, sometimes faster than an equivalent oligopeptide based on a bond that was cleaved in the protein. Legumain cleaved the bait region of rat α1-macroglobulin and was ‘trapped’ by the macroglobulin, as most other endopeptidases are, but did not interact with human α2-macroglobulin, which contains no asparagine residue in its bait region. Glycosylation of asparagine totally prevented hydrolysis by legumain. Specificity for arylamide substrates was evaluated with reference to benzyloxycarbonyl-Ala-Ala-Asn-aminomethylcoumarin, and the preference for the P3-position amino acid was Ala > Tyr(tertiary butyl) > Val > Pro > Phe = Tyr > Leu = Gly. There was no hydrolysis of substrate analogues containing mono- or di-N-methylasparagines, L-2-amino-3-ureidopropionic acid or citrulline in the P1 position. We conclude that mammalian legumain appears to be totally restricted to the hydrolysis of asparaginyl bonds in substrates of all kinds. There seem to be no strong preferences for particular amino acids in other subsites, and yet there are still unidentified factors that prevent hydrolysis of many asparaginyl bonds in proteins.

On the Structure-Function Relationships of the Unique Proteinase Inhibitor Human α2 Macroglobulin

1998 ◽  
Vol 4 (S2) ◽  
pp. 986-987
Author(s):  
James K. Stoops ◽  
Steven J. Kolodziej ◽  
Usman Qazi ◽  
Norman J. Nolasco ◽  
Peter G.W. Gettins ◽  
...  
Keyword(s):  
Structural Change ◽  
Complement Activation ◽  
Proteinase Inhibitor ◽  
Vital Role ◽  
Ester Bond ◽  
Cleavage Sites ◽  
Thiol Ester ◽  
Bait Region ◽  
Α2 Macroglobulin ◽  
Structural And Functional Properties

Human α2 macroglobulin (α2M) has structural and functional properties that contribute to its uniqueness as proteinase inhibitor. It is the largest known (Mr=720,000) and the only natural proteinase inhibitor which has a broad range of reactivity and for which the reaction is irreversible. It has a vital role in the clearance of proteinases from the circulation and in regulating their activity in fibrinolysis, coagulation and complement activation.An α2M molecule can entrap two proteinase molecules such as chymotrypsin and trypsin and can therefore be considered to contain two functional domains. Each subunit in the homotetramer has a bait region with cleavage sites for nearly all known endoproteinases and an internal thiol ester bond. A proteinase cleaves the two bait regions within both functional units leading to an activation and cleavage of the thiol ester bonds. Consequently, α2M undergoes a major structural change resulting in the irreversible entrapment of the proteinase.

scholarly journals Inhibition of intracellular proteolytic processing of soluble proproteins by an engineered α2-macroglobulin containing a furin recognition sequence in the bait region

1997 ◽  
Vol 326 (2) ◽  
pp. 507-514 ◽  
Author(s):  
Luc VAN ROMPAEY ◽  
Torik AYOUBI ◽  
Wim VAN DE VEN ◽  
Peter MARYNEN
Keyword(s):  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Proteolytic Processing ◽  
Soluble Form ◽  
Recognition Sequence ◽  
Bait Region ◽  
Α2 Macroglobulin ◽  
Ester Formation ◽  
Membrane Bound

The bait region of the general protease inhibitor α2-macroglobulin (α2M) was mutated by introducing a recognition sequence of furin. This did not interfere with folding, S-ester formation or tetramerization of the mutant recombinant α2M (rα2M). Mutant rα2M inhibited furin in vitro, by a similar mechanism to that used by plasma α2M to inhibit high-molecular-mass proteases. The mutant α2M was intracellularly active in COS-1 cells in inhibiting the endogenous processing of the soluble substrates for furin (von Willebrand factor, transforming growth factor β1 and a soluble form of the envelope glycoprotein gp160 from HIV-1) but not the membrane-bound form of gp160. The intracellular activity of mutant α2M strongly indicated that α2M attains its native conformation, and thus that the unusual internal S-ester is formed, before α2M passes through the cleavage compartment(s). Our results show for the first time that modulation of the bait region of α2M allows the creation of an inhibitor against membrane-bound proteases. It can be expected that the use of α2M-bait mutants will become important as a technique for the study of various proteolytic processes and for the identification of the proteases involved.

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