NEP_NATMA
ID NEP_NATMA Reviewed; 541 AA.
AC Q5RLZ1;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 06-FEB-2007, sequence version 2.
DT 03-AUG-2022, entry version 63.
DE RecName: Full=Halolysin-like extracellular serine protease Nep {ECO:0000305};
DE EC=3.4.21.- {ECO:0000269|PubMed:10879563};
DE AltName: Full=Subtilisin-like protease Nep {ECO:0000305};
DE Flags: Precursor;
GN Name=nep {ECO:0000303|PubMed:18553052};
OS Natrialba magadii.
OC Archaea; Euryarchaeota; Stenosarchaea group; Halobacteria; Natrialbales;
OC Natrialbaceae; Natrialba.
OX NCBI_TaxID=13769;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 122-135, SUBUNIT,
RP AND SUBCELLULAR LOCATION.
RC STRAIN=ATCC 43099;
RX PubMed=18553052; DOI=10.1007/s00792-008-0174-6;
RA De Castro R.E., Ruiz D.M., Gimenez M.I., Silveyra M.X., Paggi R.A.,
RA Maupin-Furlow J.A.;
RT "Gene cloning and heterologous synthesis of a haloalkaliphilic
RT extracellular protease of Natrialba magadii (Nep).";
RL Extremophiles 12:677-687(2008).
RN [2]
RP FUNCTION, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR
RP LOCATION, AND INDUCTION.
RC STRAIN=ATCC 43099;
RX PubMed=10879563; DOI=10.1007/s007920070033;
RA Gimenez M.I., Studdert C.A., Sanchez J.J., De Castro R.E.;
RT "Extracellular protease of Natrialba magadii: purification and biochemical
RT characterization.";
RL Extremophiles 4:181-188(2000).
RN [3]
RP INDUCTION.
RC STRAIN=ATCC 43099;
RX PubMed=20477957; DOI=10.1111/j.1472-765x.2010.02855.x;
RA Paggi R.A., Madrid E.A., D'Alessandro C.P., Cerletti M., De Castro R.E.;
RT "Growth phase-dependent biosynthesis of Nep, a halolysin-like protease
RT secreted by the alkaliphilic haloarchaeon Natrialba magadii.";
RL Lett. Appl. Microbiol. 51:36-41(2010).
RN [4]
RP FUNCTION IN PEPTIDE SYNTHESIS.
RC STRAIN=ATCC 43099;
RX PubMed=21039670; DOI=10.1111/j.1472-765x.2010.02955.x;
RA Ruiz D.M., Iannuci N.B., Cascone O., De Castro R.E.;
RT "Peptide synthesis catalysed by a haloalkaliphilic serine protease from the
RT archaeon Natrialba magadii (Nep).";
RL Lett. Appl. Microbiol. 51:691-696(2010).
RN [5]
RP EXPORT VIA THE TAT-SYSTEM, AND MUTAGENESIS OF 11-ARG-ARG-12 AND SER-351.
RC STRAIN=ATCC 43099;
RX PubMed=22582277; DOI=10.1128/jb.06792-11;
RA Ruiz D.M., Paggi R.A., Gimenez M.I., De Castro R.E.;
RT "Autocatalytic maturation of the Tat-dependent halophilic subtilase Nep
RT produced by the archaeon Natrialba magadii.";
RL J. Bacteriol. 194:3700-3707(2012).
CC -!- FUNCTION: Serine protease that hydrolyzes large proteins such as casein
CC and gelatin. Cleaves preferentially at the carboxyl terminus of Phe,
CC Tyr or Leu (PubMed:10879563). Is also able to catalyze peptide
CC synthesis under different salt concentrations in the presence of
CC dimethyl sulfoxide (DMSO) (PubMed:21039670).
CC {ECO:0000269|PubMed:10879563, ECO:0000269|PubMed:21039670}.
CC -!- ACTIVITY REGULATION: Dependent on high salt concentrations for activity
CC and stability. Strongly inhibited by the serine protease inhibitors
CC diisopropyl fluorophosphate (DFP), phenylmethyl sulfonylfluoride (PMSF)
CC and chymostatin. Also inhibited by denaturing agents such as SDS, urea,
CC and HCl guanidinium. Activated by thiol-containing reducing agents such
CC as dithiotreitol (DTT) and 2-mercaptoethanol.
CC {ECO:0000269|PubMed:10879563}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 8-10. {ECO:0000269|PubMed:10879563};
CC Temperature dependence:
CC Optimum temperature is 60 degrees Celsius.
CC {ECO:0000269|PubMed:10879563};
CC -!- SUBUNIT: Monomer. {ECO:0000303|PubMed:18553052}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:10879563,
CC ECO:0000269|PubMed:18553052}.
CC -!- INDUCTION: Expressed and secreted during the transition to the
CC stationary growth phase. Is probably up-regulated in response to
CC factors (metabolite and/or regulatory molecule) occurring in high-
CC density cultures. {ECO:0000269|PubMed:10879563,
CC ECO:0000269|PubMed:20477957}.
CC -!- PTM: Exported by the Tat system. The position of the signal peptide
CC cleavage has not been experimentally proven. After transport across the
CC membrane, the propeptide is probably processed autocatalytically,
CC yielding the mature fully active protease.
CC {ECO:0000269|PubMed:22582277}.
CC -!- SIMILARITY: Belongs to the peptidase S8 family. {ECO:0000305}.
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DR EMBL; AY804127; AAV66536.2; -; Genomic_DNA.
DR RefSeq; WP_004216473.1; NC_013922.1.
DR AlphaFoldDB; Q5RLZ1; -.
DR SMR; Q5RLZ1; -.
DR MEROPS; S08.102; -.
DR GeneID; 8823543; -.
DR OMA; WSADQGA; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:InterPro.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.200; -; 1.
DR InterPro; IPR000209; Peptidase_S8/S53_dom.
DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR InterPro; IPR022398; Peptidase_S8_His-AS.
DR InterPro; IPR023828; Peptidase_S8_Ser-AS.
DR InterPro; IPR015500; Peptidase_S8_subtilisin-rel.
DR InterPro; IPR006311; TAT_signal.
DR Pfam; PF00082; Peptidase_S8; 1.
DR PRINTS; PR00723; SUBTILISIN.
DR SUPFAM; SSF52743; SSF52743; 1.
DR PROSITE; PS51892; SUBTILASE; 1.
DR PROSITE; PS00137; SUBTILASE_HIS; 1.
DR PROSITE; PS00138; SUBTILASE_SER; 1.
DR PROSITE; PS51318; TAT; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Hydrolase; Protease; Secreted; Serine protease;
KW Signal; Zymogen.
FT SIGNAL 1..33
FT /note="Tat-type signal"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00648"
FT PROPEP 34..121
FT /evidence="ECO:0000269|PubMed:18553052"
FT /id="PRO_0000430583"
FT CHAIN 122..541
FT /note="Halolysin-like extracellular serine protease Nep"
FT /id="PRO_0000430584"
FT DOMAIN 130..405
FT /note="Peptidase S8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT REGION 403..453
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 407..430
FT /note="Acidic residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 157
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 198
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 351
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240,
FT ECO:0000303|PubMed:22582277"
FT MUTAGEN 11..12
FT /note="RR->KK: Not secreted."
FT /evidence="ECO:0000269|PubMed:22582277"
FT MUTAGEN 351
FT /note="S->A: Lack of protease activity."
FT /evidence="ECO:0000269|PubMed:22582277"
SQ SEQUENCE 541 AA; 56454 MW; 4CDC89D2F0286180 CRC64;
MTRDTNSNVG RRSVLKAASA LGAFLGLGGV ASATPGREPG PKKDEIIVGV SERVSSTEAT
VESKIPTNAE IVHTNETLGY VAVKFPSNAA EQARENFKRN VLQEDDIEYA EDNATYETLE
VPNDPMYGQQ YAPQQVNCEG AWAETYGDDD VVISVVDQGI QYDHENLAEN MDGSVSDYGY
DFVDDDGDPY PVSAGENHGT HVGGIAAGGT NNDTGHAGIS NCSMLSARAL GDGGGGSLSD
IADAIQWSAD QGADIINMSL GGGGFSETLD NACQYAYDEG TLLVAAAGND HGGSVSYPAA
YDSVMAVSSL DEGETLSSFS NVGPEIELAA PGGNVLSAVN WDDYDSLSGT SMASPVAAGV
AGLALSAHPG LSNDELRDHL HDTAVDIGLS DDEQGYGRVD AELAVTTDPD NGDDDDDDDD
DEDDPGDGEC GDETNTATAD GELSGGWGGN PSDTYSYELS TDNPCHATVT LDGPSSGATF
DLFLTLDGRT PTTSDYDRRS YNWGADEEIE VDLDGDEELG ILVDRYDGSG SYTLTIEELG
S
