AEP_YARLI
ID AEP_YARLI Reviewed; 454 AA.
AC P09230; Q6BZQ0;
DT 01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-1989, sequence version 1.
DT 03-AUG-2022, entry version 122.
DE RecName: Full=Alkaline extracellular protease {ECO:0000303|PubMed:2443483};
DE Short=AEP {ECO:0000303|PubMed:2443483};
DE EC=3.4.21.62 {ECO:0000269|PubMed:2649495, ECO:0000269|PubMed:6750031, ECO:0000269|PubMed:7007321, ECO:0000269|PubMed:870075, ECO:0000269|PubMed:9353927};
DE Flags: Precursor;
GN Name=XPR2 {ECO:0000303|PubMed:7007321}; OrderedLocusNames=YALI0F31889g;
OS Yarrowia lipolytica (strain CLIB 122 / E 150) (Yeast) (Candida lipolytica).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Dipodascaceae; Yarrowia.
OX NCBI_TaxID=284591;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2443483; DOI=10.1128/jb.169.10.4621-4629.1987;
RA Davidow L.S., O'Donnell M.M., Kaczmarek F.S., Pereira D.A., Dezeeuw J.R.,
RA Franke A.E.;
RT "Cloning and sequencing of the alkaline extracellular protease gene of
RT Yarrowia lipolytica.";
RL J. Bacteriol. 169:4621-4629(1987).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-200, SUBCELLULAR LOCATION,
RP GLYCOSYLATION AT ASN-123, AND PROTEOLYTIC PROCESSING.
RX PubMed=3211132; DOI=10.1128/mcb.8.11.4904-4916.1988;
RA Matoba S., Fukayama J., Wing R.A., Ogrydziak D.M.;
RT "Intracellular precursors and secretion of alkaline extracellular protease
RT of Yarrowia lipolytica.";
RL Mol. Cell. Biol. 8:4904-4916(1988).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA] OF 35-454.
RC STRAIN=CLIB 122 / E 150;
RX PubMed=15229592; DOI=10.1038/nature02579;
RA Dujon B., Sherman D., Fischer G., Durrens P., Casaregola S., Lafontaine I.,
RA de Montigny J., Marck C., Neuveglise C., Talla E., Goffard N., Frangeul L.,
RA Aigle M., Anthouard V., Babour A., Barbe V., Barnay S., Blanchin S.,
RA Beckerich J.-M., Beyne E., Bleykasten C., Boisrame A., Boyer J.,
RA Cattolico L., Confanioleri F., de Daruvar A., Despons L., Fabre E.,
RA Fairhead C., Ferry-Dumazet H., Groppi A., Hantraye F., Hennequin C.,
RA Jauniaux N., Joyet P., Kachouri R., Kerrest A., Koszul R., Lemaire M.,
RA Lesur I., Ma L., Muller H., Nicaud J.-M., Nikolski M., Oztas S.,
RA Ozier-Kalogeropoulos O., Pellenz S., Potier S., Richard G.-F.,
RA Straub M.-L., Suleau A., Swennen D., Tekaia F., Wesolowski-Louvel M.,
RA Westhof E., Wirth B., Zeniou-Meyer M., Zivanovic Y., Bolotin-Fukuhara M.,
RA Thierry A., Bouchier C., Caudron B., Scarpelli C., Gaillardin C.,
RA Weissenbach J., Wincker P., Souciet J.-L.;
RT "Genome evolution in yeasts.";
RL Nature 430:35-44(2004).
RN [4]
RP PROTEIN SEQUENCE OF 158-182, SUBCELLULAR LOCATION, FUNCTION, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, AND
RP GLYCOSYLATION.
RX PubMed=6750031; DOI=10.1099/00221287-128-6-1225;
RA Ogrydziak D.M., Scharf S.J.;
RT "Alkaline extracellular protease produced by Saccharomycopsis lipolytica
RT CX161-1B.";
RL J. Gen. Microbiol. 128:1225-1234(1982).
RN [5]
RP INDUCTION, FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=870075; DOI=10.1016/0304-4165(77)90209-4;
RA Ogrydziak D.M., Demain A.L., Tannenbaum S.R.;
RT "Regulation of extracellular protease production in Candida lipolytica.";
RL Biochim. Biophys. Acta 497:525-538(1977).
RN [6]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=7007321; DOI=10.1128/jb.145.1.404-409.1981;
RA Simms P.C., Ogrydziak D.M.;
RT "Structural gene for the alkaline extracellular protease of
RT Saccharomycopsis lipolytica.";
RL J. Bacteriol. 145:404-409(1981).
RN [7]
RP PROTEOLYTIC PROCESSING, ACTIVITY REGULATION, FUNCTION, AND CATALYTIC
RP ACTIVITY.
RX PubMed=2649495; DOI=10.1016/s0021-9258(18)83309-9;
RA Matoba S., Ogrydziak D.M.;
RT "A novel location for dipeptidyl aminopeptidase processing sites in the
RT alkaline extracellular protease of Yarrowia lipolytica.";
RL J. Biol. Chem. 264:6037-6043(1989).
RN [8]
RP PROTEOLYTIC PROCESSING, FUNCTION OF THE PRO-REGION, SUBCELLULAR LOCATION,
RP AND GLYCOSYLATION.
RX PubMed=1995632; DOI=10.1016/s0021-9258(19)67863-4;
RA Fabre E., Nicaud J.-M., Lopez M.C., Gaillardin C.;
RT "Role of the proregion in the production and secretion of the Yarrowia
RT lipolytica alkaline extracellular protease.";
RL J. Biol. Chem. 266:3782-3790(1991).
RN [9]
RP PROTEOLYTIC PROCESSING, FUNCTION OF THE PRO-REGION, AND SUBCELLULAR
RP LOCATION.
RX PubMed=1634541; DOI=10.1016/s0021-9258(18)42144-8;
RA Fabre E., Tharaud C., Gaillardin C.;
RT "Intracellular transit of a yeast protease is rescued by trans-
RT complementation with its prodomain.";
RL J. Biol. Chem. 267:15049-15055(1992).
RN [10]
RP PROTEOLYTIC PROCESSING, SUBCELLULAR LOCATION, AND SECRETION PATHWAY
RP OVERLOAD.
RX PubMed=7954894; DOI=10.1007/bf00326302;
RA Le Dall M.T., Nicaud J.M., Gaillardin C.;
RT "Multiple-copy integration in the yeast Yarrowia lipolytica.";
RL Curr. Genet. 26:38-44(1994).
RN [11]
RP INDUCTION.
RX PubMed=8264600; DOI=10.1128/mcb.14.1.327-338.1994;
RA Blanchin-Roland S., Cordero Otero R.R., Gaillardin C.;
RT "Two upstream activation sequences control the expression of the XPR2 gene
RT in the yeast Yarrowia lipolytica.";
RL Mol. Cell. Biol. 14:327-338(1994).
RN [12]
RP PROTEOLYTIC PROCESSING BY XPR6.
RX PubMed=8203153; DOI=10.1002/yea.320100107;
RA Enderlin C.S., Ogrydziak D.M.;
RT "Cloning, nucleotide sequence and functions of XPR6, which codes for a
RT dibasic processing endoprotease from the yeast Yarrowia lipolytica.";
RL Yeast 10:67-79(1994).
RN [13]
RP SUBCELLULAR LOCATION.
RX PubMed=8798620; DOI=10.1074/jbc.271.39.23895;
RA Mamoun C.B., Beckerich J.M., Gaillardin C.;
RT "The TSR1 gene of Yarrowia lipolytica is involved in the signal recognition
RT particle-dependent translocation pathway of secretory proteins.";
RL J. Biol. Chem. 271:23895-23901(1996).
RN [14]
RP INDUCTION.
RX PubMed=8842151; DOI=10.1007/bf02173777;
RA Otero R.C., Gaillardin C.;
RT "Dominant mutations affecting expression of pH-regulated genes in Yarrowia
RT lipolytica.";
RL Mol. Gen. Genet. 252:311-319(1996).
RN [15]
RP INDUCTION.
RX PubMed=9308186; DOI=10.1099/00221287-143-9-3045;
RA Glover D.J., McEwen R.K., Thomas C.R., Young T.W.;
RT "pH-regulated expression of the acid and alkaline extracellular proteases
RT of Yarrowia lipolytica.";
RL Microbiology 143:3045-3054(1997).
RN [16]
RP SIGNAL PEPTIDE, PROTEOLYTIC PROCESSING, SUBCELLULAR LOCATION, MUTAGENESIS
RP OF SER-397, FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=9353927; DOI=10.1099/00221287-143-10-3263;
RA Matoba S., Morano K.A., Klionsky D.J., Kim K., Ogrydziak D.M.;
RT "Dipeptidyl aminopeptidase processing and biosynthesis of alkaline
RT extracellular protease from Yarrowia lipolytica.";
RL Microbiology 143:3263-3272(1997).
RN [17]
RP INDUCTION.
RX PubMed=9199331; DOI=10.1128/mcb.17.7.3966;
RA Lambert M., Blanchin-Roland S., Le Louedec F., Lepingle A., Gaillardin C.;
RT "Genetic analysis of regulatory mutants affecting synthesis of
RT extracellular proteinases in the yeast Yarrowia lipolytica: identification
RT of a RIM101/pacC homolog.";
RL Mol. Cell. Biol. 17:3966-3976(1997).
RN [18]
RP INDUCTION.
RX PubMed=10206713; DOI=10.1099/13500872-145-1-75;
RA Madzak C., Blanchin-Roland S., Cordero-Otero R.R., Gaillardin C.;
RT "Functional analysis of upstream regulating regions from the Yarrowia
RT lipolytica XPR2 promoter.";
RL Microbiology 145:75-87(1999).
RN [19]
RP INDUCTION.
RX PubMed=11861549; DOI=10.1093/genetics/160.2.417;
RA Gonzalez-Lopez C.I., Szabo R., Blanchin-Roland S., Gaillardin C.;
RT "Genetic control of extracellular protease synthesis in the yeast Yarrowia
RT lipolytica.";
RL Genetics 160:417-427(2002).
RN [20]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=17432872; DOI=10.1021/jf0633894;
RA Poza M., Sestelo A.B., Ageitos J.M., Vallejo J.A., Veiga-Crespo P.,
RA Villa T.G.;
RT "Cloning and expression of the XPR2 gene from Yarrowia lipolytica in Pichia
RT pastoris.";
RL J. Agric. Food Chem. 55:3944-3948(2007).
RN [21]
RP PROTEOLYTIC PROCESSING, SUBCELLULAR LOCATION, AND SECRETION PATHWAY
RP OVERLOAD.
RX PubMed=22909173; DOI=10.1111/j.1567-1364.2012.00846.x;
RA Ogrydziak D.M., Nicaud J.M.;
RT "Characterization of Yarrowia lipolytica XPR2 multi-copy strains over-
RT producing alkaline extracellular protease - a system for rapidly increasing
RT secretory pathway cargo loads.";
RL FEMS Yeast Res. 12:938-948(2012).
CC -!- FUNCTION: Major secreted protein that belongs to the subtilisin family
CC serine proteases. {ECO:0000269|PubMed:17432872,
CC ECO:0000269|PubMed:2649495, ECO:0000269|PubMed:6750031,
CC ECO:0000269|PubMed:7007321, ECO:0000269|PubMed:870075,
CC ECO:0000269|PubMed:9353927}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of proteins with broad specificity for peptide
CC bonds, and a preference for a large uncharged residue in P1.
CC Hydrolyzes peptide amides.; EC=3.4.21.62;
CC Evidence={ECO:0000269|PubMed:17432872, ECO:0000269|PubMed:2649495,
CC ECO:0000269|PubMed:6750031, ECO:0000269|PubMed:7007321,
CC ECO:0000269|PubMed:870075, ECO:0000269|PubMed:9353927};
CC -!- ACTIVITY REGULATION: The protease activity is completely inhibited by
CC the serine inhibitor PMSF but is not affected by thiol group inhibitors
CC and in the presence of dithiothreitol (PubMed:6750031). In the presence
CC of high concentrations of o-phenanthroline the protease activity is
CC only partially inhibited (PubMed:6750031). The pro-region plays an
CC inhibitory role and may provide a mechanism for preventing premature
CC activation in the secretory pathway (PubMed:2649495).
CC {ECO:0000269|PubMed:2649495, ECO:0000269|PubMed:6750031}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 9.0-10.0. {ECO:0000269|PubMed:17432872,
CC ECO:0000269|PubMed:6750031};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:6750031};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:1634541,
CC ECO:0000269|PubMed:1995632, ECO:0000269|PubMed:22909173,
CC ECO:0000269|PubMed:3211132, ECO:0000269|PubMed:6750031,
CC ECO:0000269|PubMed:7954894, ECO:0000269|PubMed:8798620,
CC ECO:0000269|PubMed:9353927}. Note=Proper secretion requires TSR1.
CC {ECO:0000269|PubMed:8798620}.
CC -!- INDUCTION: Expression is subject to at least 3 different regulatory
CC controls, carbon, sulfur and nitrogen repression (PubMed:870075).
CC Intracellular cysteine and ammonia appear to be the metabolic signals
CC for sulfur and nitrogen repression (PubMed:870075). Moreover, pH
CC regulates expression independently from other metabolic signals, with
CC highest levels of AEP mRNA at pH 6.5 (PubMed:8842151, PubMed:9308186).
CC The transcriptional activator RIM101 and the Rim pathway are required
CC for the alkaline induction of gene expression (PubMed:9199331,
CC PubMed:11861549). Two major upstream activation sequences (UASs) are
CC essential for promoter activity under conditions of repression or full
CC induction. The distal UAS (UAS1) is located at position -790 to -778,
CC whereas the proximal UAS (UAS2) localizes at positions -148 to -124
CC (PubMed:8264600, PubMed:10206713). {ECO:0000269|PubMed:10206713,
CC ECO:0000269|PubMed:11861549, ECO:0000269|PubMed:8264600,
CC ECO:0000269|PubMed:870075, ECO:0000269|PubMed:8842151,
CC ECO:0000269|PubMed:9199331, ECO:0000269|PubMed:9308186}.
CC -!- PTM: The pro-region is removed through cleavage by XPR6 after Lys156-
CC Arg157, which yields mature active XPR2. {ECO:0000269|PubMed:1634541,
CC ECO:0000269|PubMed:22909173, ECO:0000269|PubMed:3211132,
CC ECO:0000269|PubMed:7954894, ECO:0000269|PubMed:8203153,
CC ECO:0000269|PubMed:9353927}.
CC -!- PTM: The 10 consecutive -X-Ala- or -X-Pro- dipeptides located over 100
CC amino acids upstream of the N-terminal of mature XPR2 are subject to
CC dipeptidyl aminopeptidase (DPAPase)-processing (PubMed:9353927).
CC DPAPase activity is not necessary for XPR6 cleavage and for secretion
CC of mature active XPR2 (PubMed:9353927). {ECO:0000269|PubMed:1995632,
CC ECO:0000269|PubMed:2649495, ECO:0000269|PubMed:8203153,
CC ECO:0000269|PubMed:9353927}.
CC -!- PTM: N-glycosylated. Glycosylation within the pro-region has no effect
CC on secretion and maturation at 18 degrees Celsius, but is required for
CC secretion at 28 degrees Celsius (PubMed:1995632).
CC {ECO:0000269|PubMed:1995632, ECO:0000269|PubMed:3211132,
CC ECO:0000269|PubMed:6750031}.
CC -!- MISCELLANEOUS: The pro-region inhibits protease activity
CC (PubMed:2649495) and plays an additional essential role in the proper
CC folding of the protein into a conformation compatible with secretion
CC (PubMed:1995632, PubMed:1634541). {ECO:0000269|PubMed:1634541,
CC ECO:0000269|PubMed:1995632, ECO:0000269|PubMed:2649495}.
CC -!- MISCELLANEOUS: Its complex processing and high level of secretion make
CC XPR2 the perfect model to study the secretion pathway.
CC {ECO:0000269|PubMed:22909173, ECO:0000269|PubMed:7954894}.
CC -!- SIMILARITY: Belongs to the peptidase S8 family. {ECO:0000305}.
CC -!- CAUTION: Strain CLIB 122 / E 150 has a defective XPR2 sequence (xpr2-
CC 322) which lacks the N-terminus (positions 1 to 34). {ECO:0000305}.
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DR EMBL; M17741; AAA35242.1; -; Genomic_DNA.
DR EMBL; M23353; AAA35250.1; -; Genomic_DNA.
DR EMBL; CR382132; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR PIR; A26955; A26955.
DR AlphaFoldDB; P09230; -.
DR SMR; P09230; -.
DR MEROPS; S08.055; -.
DR iPTMnet; P09230; -.
DR InParanoid; P09230; -.
DR Proteomes; UP000001300; Chromosome F.
DR GO; GO:0005615; C:extracellular space; IBA:GO_Central.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IBA:GO_Central.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR CDD; cd04077; Peptidases_S8_PCSK9_ProteinaseK_like; 1.
DR Gene3D; 3.40.50.200; -; 1.
DR InterPro; IPR034193; PCSK9_ProteinaseK-like.
DR InterPro; IPR000209; Peptidase_S8/S53_dom.
DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR InterPro; IPR023827; Peptidase_S8_Asp-AS.
DR InterPro; IPR022398; Peptidase_S8_His-AS.
DR InterPro; IPR023828; Peptidase_S8_Ser-AS.
DR InterPro; IPR015500; Peptidase_S8_subtilisin-rel.
DR InterPro; IPR010259; S8pro/Inhibitor_I9.
DR Pfam; PF05922; Inhibitor_I9; 1.
DR Pfam; PF00082; Peptidase_S8; 1.
DR PRINTS; PR00723; SUBTILISIN.
DR SUPFAM; SSF52743; SSF52743; 1.
DR PROSITE; PS51892; SUBTILASE; 1.
DR PROSITE; PS00136; SUBTILASE_ASP; 1.
DR PROSITE; PS00137; SUBTILASE_HIS; 1.
DR PROSITE; PS00138; SUBTILASE_SER; 1.
PE 1: Evidence at protein level;
KW Cleavage on pair of basic residues; Direct protein sequencing;
KW Glycoprotein; Hydrolase; Protease; Reference proteome; Secreted;
KW Serine protease; Signal; Zymogen.
FT SIGNAL 1..15
FT /evidence="ECO:0000269|PubMed:9353927"
FT PROPEP 16..157
FT /evidence="ECO:0000269|PubMed:6750031,
FT ECO:0000269|PubMed:9353927"
FT /id="PRO_0000026984"
FT CHAIN 158..454
FT /note="Alkaline extracellular protease"
FT /id="PRO_0000026985"
FT DOMAIN 68..146
FT /note="Inhibitor I9"
FT /evidence="ECO:0000255"
FT DOMAIN 166..454
FT /note="Peptidase S8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 200
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 231
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 397
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT CARBOHYD 123
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:3211132"
FT MUTAGEN 397
FT /note="S->A: Abolishes protease activity, but not
FT maturation."
FT /evidence="ECO:0000269|PubMed:9353927"
SQ SEQUENCE 454 AA; 46906 MW; 3DFBA196048B191E CRC64;
MKLATAFTIL TAVLAAPLAA PAPAPDAAPA AVPEGPAAAA YSSILSVVAK QSKKFKHHKR
DLDEKDQFIV VFDSSATVDQ IASEIQKLDS LVDEDSSNGI TSALDLPVYT DGSGFLGFVG
KFNSTIVDKL KESSVLTVEP DTIVSLPEIP ASSNAKRAIQ TTPVTQWGLS RISHKKAQTG
NYAYVRETVG KHPTVSYVVD SGIRTTHSEF GGRAVWGANF ADTQNADLLG HGTHVAGTVG
GKTYGVDANT KLVAVKVFAG RSAALSVINQ GFTWALNDYI SKRDTLPRGV LNFSGGGPKS
ASQDALWSRA TQEGLLVAIA AGNDAVDACN DSPGNIGGST SGIITVGSID SSDKISVWSG
GQGSNYGTCV DVFAPGSDII SASYQSDSGT LVYSGTSMAC PHVAGLASYY LSINDEVLTP
AQVEALITES NTGVLPTTNL KGSPNAVAYN GVGI
