3.4.21.92: Endopeptidase Clp

This is an abbreviated version!
For detailed information about Endopeptidase Clp, go to the full flat file.

Word Map on EC 3.4.21.92

Reaction

Hydrolysis of proteins to small peptides in the presence of ATP and Mg2+. alpha-Casein is the usual test substrate. In the absence of ATP, only oligopeptides shorter than five residues are hydrolysed (such as succinyl-Leu-Tyr-/-NHMec; and Leu-Tyr-Leu-/-Tyr-Trp, in which cleavage of the -Tyr-/-Leu- and -Tyr-/-Trp bonds also occurs) =

Synonyms

Protease Ti, ATP-dependent Clp protease, endopeptidase Ti, Caseinolytic protease, ClpP, Clp protease, endopeptidase Clp, Heat shock protein F21.5, stress protein G7, ClpQ, ClpC, ClpA, ClpB, ClpX, ClpY, ClpAP protease, ClpS1, nClpP7, nClpP8, ClpP protease complex, CLP, ClpXP, ClpCP protease, ClpC ATPase, ClpP Protease, ClpP Peptidase, Clp proteolytic subunit, heat-shock protease ClpP, ClpAP, ClpP3, ClpR, ClpP3/R complex, ClpCP3/R protease, ClpE, PfClpP, ClpX2, ATP-dependent caseinolytic protease, ClpP1 protease, ClpP2 protease, ClpXP protease, BsClpP, ClpC1, ATP-dependent Clp protease proteolytic subunit 1, ClpP1, ATP-dependent Clp protease proteolytic subunit 2, ClpP2, CplC, ClpP1P2

ECTree

     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.21 Serine endopeptidases
                3.4.21.92 Endopeptidase Clp

Engineering

Engineering on EC 3.4.21.92 - Endopeptidase Clp

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G773R
-
inactive mutant
K229A
-
RAH mutant to investigate the role of the RKH sequence loops
DELTA1-17
-
deletion of the N-terminal 10, 14, or 17 residues of mature ClpP allows these mutants to degrade alpha-casein, a natively unfolded protein
K25A
-
mutant shows wild-type level of dipeptide cleavage, 1.5fold increase in decapeptide cleavage compared to wild-type. Mutant shows only moderate decrease in ClpX affinity
S499C
-
mutation introduced for FRET measurements. Labelling of S499C with fluorescent probes induces a 10-20fold increase in both chaperone and ATPase activities
R22A
-
mutant shows wild-type level of dipeptide cleavage, 1.5fold increase in decapeptide cleavage compared to wild-type. Mutant shows high decrease in ClpX affinity
S21A
-
mutant shows wild-type level of dipeptide cleavage, 5fold increase in decapeptide cleavage compared to wild-type. Mutant shows high decrease in ClpX affinity
Delta1-10
-
deletion of the N-terminal 10, 14, or 17 residues of mature ClpP allows these mutants to degrade alpha-casein, a natively unfolded protein
I19A
-
mutant shows wild-type level of dipeptide cleavage, 20fold increase in decapeptide cleavage compared to wild-type, in contrast to wild-type mutant degrades 113-residue unfolded I27 domain of human titin. Mutant shows high decrease in ClpX affinity
L23A
-
mutant shows wild-type level of dipeptide cleavage, 5fold increase in decapeptide cleavage compared to wild-type. Mutant shows only moderate decrease in ClpX affinity
H230A
-
RKH mutant to investigate the role of the RKH sequence loops
R228A
-
AKH mutant to investigate the role of the RKH sequence loops
E8G/Q9G/T10G/S11G/R12G/G13G/E14G/R15G
-
residues 8-15 form the channel loop of the pore: when replaced with eight glycines this mutant cleaves the decapeptide at a rate 8fold faster than observed with wild-type ClpP but cleaves the dipeptide at a comparable rate. Mutant shows a much slower degradation of GFP-ssrA
E8R/R12E/E14R/R15E
-
residues 8-15 form the channel loop of the pore: when charged residues in the channel (amino acids 8-15) are reversed this mutant cleaves the decapeptide at a rate 8fold faster than observed with wild-type ClpP but cleaves the dipeptide at a comparable rate. Mutant shows a much slower degradation of GFP-ssrA
E8G/R12G/E14G/R15G
-
residues 8-15 form the channel loop of the pore: when charged residues in the channel (amino acids 8-15) are changed with glycine this mutant cleaves the decapeptide at a rate 8fold faster than observed with wild-type ClpP but cleaves the dipeptide at a comparable rate. Mutant shows a much slower degradation of GFP-ssrA
E8A/R12A/E14A/R15A
-
residues 8-15 form the channel loop of the pore: when charged residues in the channel (amino acids 8-15) are changed with alanine this mutant cleaves the decapeptide at a rate 8fold faster than observed with wild-type ClpP but cleaves the dipeptide at a comparable rate. Mutant shows a substantial GFP-ssrA degradation similar to wild-type
Y20A
-
mutant shows wild-type level of dipeptide cleavage, 5fold increase in decapeptide cleavage compared to wild-type. Mutant shows high decrease in ClpX affinity
L24A
-
mutant shows wild-type level of dipeptide cleavage, 20fold increase in decapeptide cleavage compared to wild-type. Mutant shows high decrease in ClpX affinity
I19L
-
mutant shows 6fold increase in decapeptide cleavage compared to wild-type
I19D
-
mutant shows 16fold increase in decapeptide cleavage compared to wild-type
DELTA1-14
-
deletion of the N-terminal 10, 14, or 17 residues of mature ClpP allows these mutants to degrade alpha-casein, a natively unfolded protein
A153C
-
the structure of a crosslinked Escherichia coli ClpP are determined in which the two heptameric rings of ClpP are held together by disulfide bonds. While all Escherichia coli ClpP structures solved to date are in the extended state, the crosslinked ClpP structure is found to be in the compact state. Under reducing condition Km (N-succinyl-Leu-Tyr-7-amido-4-methylcoumarin) is similar to wild-type but kcat is 32fold lower. Under non-reducing conditions mutant is inactive and does not bind its cognate chaperone
S499C
-
mutation introduced for FRET measurements. Labelling of S499C with fluorescent probes induces a 10-20fold increase in both chaperone and ATPase activities
-
S99A
-
does not digest peptide into smaller fragments
S97A
inactive. Overexpression has no effect on sensitivity of cells to cisplatin
S61A/Y63V/L83A/Y91V
mutations in hydrophobic patch of subunit ClpP1. Complex formation and processing still occurs. The complex containing the mutant is catalytically active
Y75V/Y95V
mutations in hydrophobic patch of subunit ClpP2.Complex formation and processing still occurs. The complex containing the mutant is catalytically inactive
S98A
-
mutant shows no activity
Y63A
gain-of-function mutant, inhibits staphylococcal growth. The mutant becomes an uncontrolled protease that actively hydrolyzes cell division protein FtsZ in vitro. Expression of mutant phenocopies treatment with acyldepsipeptides, inhibition of cell division occurs as does showing sterilizing with rifampicin antibiotics
G127
-
mutant shows no activity
G131
-
mutant shows no activity
G128
-
mutant shows no activity
E135R
-
mutant shows no activity
E135A
-
mutant shows no activity
L144M
-
Km (N-succinyl-Leu-Tyr-7-amido-4-methylcoumarin): 0.523 mM, kcat: 0.003/sec
L144R
-
mutant shows no activity
L144E
-
mutant shows no activity
L144G
-
mutant shows no activity
Y63A
-
gain-of-function mutant, inhibits staphylococcal growth. The mutant becomes an uncontrolled protease that actively hydrolyzes cell division protein FtsZ in vitro. Expression of mutant phenocopies treatment with acyldepsipeptides, inhibition of cell division occurs as does showing sterilizing with rifampicin antibiotics
-
A153P
-
crystallization data, disruption of handle region resulting in an altered ring-ring dimerization interface
S101A
-
mutation of the active site Ser-101 in ClpP3 inactivates the entire ClpCP3/R protease, the mutation has no effect on the formation of the recombinant ClpP3/R core complex of 270 kDa. The mutated ClpP3/R complex stimulates the steady-state ATPase activity of ClpC to the same extent as wild-type ClpP3/R. The mutated ClpP3/R core fails to degrade the alpha-casein nor the more sensitive FITC-casein
additional information