The proteolytic specificity of the natural enediyne-containing chromoproteins is unique to each chromoprotein

Zein, Nada; Reiss, Paul D.; Bernatowicz, Michael S.; Bolgar, Mark S.

doi:10.1016/1074-5521(95)90262-7

Cited by 46 publications

(25 citation statements)

References 26 publications

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“…4) within 2 h of incubation. This result is similar to those previously reported (23,24). However, a noticeable difference is observed in the proteolytic activity of the recombinant and natural proteins, because although the concentration of the recombinant protein was higher, the time required to obtain full proteolysis of the sample was 10 times longer than for natural NCS.…”

Section: Resultssupporting

confidence: 79%

“…This would also explain why proteins with similar structures and sequences are associated with various proteolytic activities, from exopeptidase to endopeptidases (18,21,22,24). The activity being due to a contaminant would also account for the very low specific activity: the experimental conditions used (apoprotein/substrate ratio, incubation time) indicate a specific activity at least 3 orders of magnitude lower than that of typical proteases like trypsin.…”

Section: Discussionmentioning

confidence: 99%

“…Each chromoprotein seems to have a characteristic proteolytic specificity (22). Whereas C-1027 (18) and macromomycin (21) seem to have aminopeptidase activity, endopeptidase activity has been shown for kedarcidin (18), maduropeptin, and NCS (23,24).…”

mentioning

confidence: 99%

“…These activities have been detected with either histones (18,(21)(22)(23)(24) or synthetic peptides (24). Histones were chosen as substrates because it was thought that these highly basic proteins would probably interact with the acid-rich apoproteins.…”

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confidence: 99%

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Reinvestigation of the Proteolytic Activity of Neocarzinostatin

et al. 2000

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Neocarzinostatin (NCS) is the most studied member of a family of chromoproteins secreted by a range of actinomycetes species. It has been proposed that in addition to their antitumoral activity related to the bound chromophores, this group of related proteins could be a secreted proteases superfamily. With the aim of dissecting the molecular basis of the proteolytic activity of NCS, an expression system allowing efficient expression of apo-NCS in Escherichia coli was constructed. The recombinant protein was properly folded and functional. Its histone-specific proteolytic activity was similar to the activity described for the natural protein.Further analyses unambiguously demonstrated that the proteolytic activity could be physically separated from NCS. This activity is therefore due not to NCS itself but to minor contaminating proteases, the nature of which differed in the recombinant and natural NCS preparations. The histone degradation test commonly used to monitor proteolytic activity is extremely sensitive and may easily generate false-positive results. These results strongly suggest that the possible proteolytic activity of the proteins of this family should be critically reconsidered.

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Section: Resultssupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Reinvestigation of the Proteolytic Activity of Neocarzinostatin

et al. 2000

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“…Given in parentheses are nucleotide sequence accession numbers for cagA, axnA, ncsA, and mcmA, and protein sequence accession number for Ked. then cut by the chromophore (36,50). In addition, the observation that the expression of cagA and mcmA is constitutive and independent of the chromophore production, assuring that the chromophore will be sequestered, suggests the apoprotein may function in a self-resistance role as well (43,45).…”

Section: Figmentioning

confidence: 95%