Expression of the pro-regions of papain and papaya proteinase IV in <i>Escherichia coli</i> and their inhibition of mature cysteine proteinases

Taylor, Mark A.; Briggs, Geoffrey S.; Baker, Kenneth C.; Cummings, Nicola J.; Pratt, Kathryn A.; Freedman, Robert B.; Goodenough, Peter W.

doi:10.1042/bst023080s

Cited by 10 publications

(4 citation statements)

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“…Similar results have been published for members of the cathepsin L‐like subfamily, e.g. papain and cathepsins L and S [9–12]. In each case, the propeptide binds to the cognate protease with high affinity, albeit not with absolute specificity.…”

Section: Introductionsupporting

confidence: 81%

The α1/2 helical backbone of the prodomains defines the intrinsic inhibitory specificity in the cathepsin L‐like cysteine protease subfamily

et al. 2000

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Proregions of papain-like cysteine proteases are potent and often highly selective inhibitors of their parental enzymes. The molecular basis of their selectivity is poorly understood. For two closely related members of the cathepsin Llike subfamily we established strong selectivity differences. The propeptide of cathepsin S was observed to inhibit cathepsin L with a K i of 0.08 nM, yet cathepsin L propeptide inhibited cathepsin S only poorly. To identify the respective structural correlates we engineered chimeric propeptides and compared their inhibitory specificity with the wild-types. Specificity resided in the N-terminal part, strongly suggesting that the backbone of the prodomain was the underlying structure.z 2000 Federation of European Biochemical Societies.

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Section: Introductionsupporting

confidence: 81%

The α1/2 helical backbone of the prodomains defines the intrinsic inhibitory specificity in the cathepsin L‐like cysteine protease subfamily

et al. 2000

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“…In fact the prosegment has been shown to be a potent and specific inhibitor of its associated protease. This has been demonstrated with cathepsins B and L [3,4], papain [5], subtilisin [6,7], pepsin [8] and stromelysin [9]. This unique property of the prosegment can thus be seen as an additional strategy to regulate enzymic activity in delaying its onset until the moment when the enzyme needs to be fully functional.…”

Section: Introductionmentioning

confidence: 99%

Synthetic peptides derived from the prosegments of proprotein convertase 1/3 and furin are potent inhibitors of both enzymes

Basak

Lazure

2003

Biochemical Journal

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Proprotein convertases (PCs) are Ca(2+)-dependent serine proteases of the subtilisin/kexin family which are known specifically to cleave propeptide and proprotein substrates at the C-terminal of R-X-(K/R)-R/ to generate the relevant biologically active peptides. PCs are initially synthesized as enzymically inactive proenzyme forms where the prosegments play an important inhibitory role to the respective enzymes. Here we investigated whether synthetic peptides derived from the pro-region could also represent specific and potent inhibitors. Based upon sequence alignment, secondary structure analysis and hydrophilicity plot, a number of peptides ranging from 8 to 33 residues were selected. These included segments encompassing residues 55-62, 50-62, 39-62, 50-83, 55-83, 64-83 and 74-83 in the pro-mouse PC1/3 sequence and residues 54-62, 48-62 and 39-62 of the pro-human furin sequence. All peptides were prepared by solid-phase FastMoc chemistry, purified by reversed-phase HPLC and characterized by MS and amino acid analysis. These peptides were tested in vitro for inhibitory activity towards recombinant mouse PC1/3 and human furin. Progress-curve and end-time kinetic analysis demonstrated that a number of these peptides, particularly those containing both the primary and the secondary processing sites, displayed strong inhibition of both enzymes with inhibition constants (K (i)) in the high nanomolar range. Unlike the whole propeptide, these small synthetic peptide inhibitors exhibited either true competitive or mixed competitive inhibition, depending on the sequence. Our data revealed further the critical role of the last two basic amino acid residues (e.g. Lys(82)-Arg(83) for the mouse PC1/3 sequence) of the prodomain in imparting a strong anti-convertase activity. The study also establishes the inhibitory potential of certain regions contained within the prosegment of the two convertases.

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“…It is important to note that eventual translation of the GMCP3« mRNA would result in a truncated protein containing the whole propeptide. Given the inhibitory activity described for this domain (Taylor et al, 1995;Chagas et al, 1996), it is tempting to speculate a possible function of the GMCP3 paralogue to trans-regulate the protease activity of GMCP3.…”

Section: Discussionmentioning

confidence: 99%

A distinct subfamily of papain-like cystein proteinases regulated by senescence and stresses in Glycine max

Esteban-García

Garrido-Cárdenas

Alonso

et al. 2010

Journal of Plant Physiology

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Expression of the pro-regions of papain and papaya proteinase IV in Escherichia coli and their inhibition of mature cysteine proteinases

Cited by 10 publications

References 0 publications

The α1/2 helical backbone of the prodomains defines the intrinsic inhibitory specificity in the cathepsin L‐like cysteine protease subfamily

The α1/2 helical backbone of the prodomains defines the intrinsic inhibitory specificity in the cathepsin L‐like cysteine protease subfamily

Synthetic peptides derived from the prosegments of proprotein convertase 1/3 and furin are potent inhibitors of both enzymes

A distinct subfamily of papain-like cystein proteinases regulated by senescence and stresses in Glycine max

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