IBP_LEUSY
ID IBP_LEUSY Reviewed; 261 AA.
AC C7F6X3;
DT 18-JUL-2018, integrated into UniProtKB/Swiss-Prot.
DT 22-SEP-2009, sequence version 1.
DT 25-MAY-2022, entry version 40.
DE RecName: Full=Ice-binding protein {ECO:0000303|PubMed:20067781};
DE AltName: Full=Antifreeze protein {ECO:0000312|EMBL:ACU30806.1};
DE Short=AFP {ECO:0000305};
DE AltName: Full=LeIBP {ECO:0000303|PubMed:21795798};
DE Flags: Precursor;
GN Name=AFP {ECO:0000312|EMBL:ACU30806.1};
OS Leucosporidium sp. (strain AY30) (Arctic yeast).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Pucciniomycotina;
OC Microbotryomycetes; Leucosporidiales; Leucosporidium;
OC unclassified Leucosporidium.
OX NCBI_TaxID=662878;
RN [1] {ECO:0000312|EMBL:ACU30806.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=AY30 {ECO:0000312|EMBL:ACU30806.1};
RA Lee J.K., Kang S.H., Kim H.J.;
RT "Gene cloning of antifreeze protein (AFP) of Leucosporidium sp. AY30.";
RL Submitted (JUN-2009) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 214-226, IDENTIFICATION BY
RP MASS SPECTROMETRY, FUNCTION, SUBCELLULAR LOCATION, AND GLYCOSYLATION.
RX PubMed=20067781; DOI=10.1016/j.cryobiol.2010.01.002;
RA Lee J.K., Park K.S., Park S., Park H., Song Y.H., Kang S.H., Kim H.J.;
RT "An extracellular ice-binding glycoprotein from an Arctic psychrophilic
RT yeast.";
RL Cryobiology 60:222-228(2010).
RN [3]
RP PROTEIN SEQUENCE OF 21-29, FUNCTION, SUBUNIT, SUBCELLULAR LOCATION,
RP GLYCOSYLATION, SIGNAL SEQUENCE, AND CIRCULAR DICHROISM ANALYSIS.
RX PubMed=22426061; DOI=10.1016/j.cryobiol.2012.02.014;
RA Park K.S., Do H., Lee J.H., Park S.I., Kim E., Kim S.J., Kang S.H.,
RA Kim H.J.;
RT "Characterization of the ice-binding protein from Arctic yeast
RT Leucosporidium sp. AY30.";
RL Cryobiology 64:286-296(2012).
RN [4]
RP PROTEIN SEQUENCE OF 21-25, FUNCTION, SUBCELLULAR LOCATION, GLYCOSYLATION,
RP MASS SPECTROMETRY, AND CIRCULAR DICHROISM ANALYSIS.
RX PubMed=23203635; DOI=10.1007/s00253-012-4594-y;
RA Lee J.H., Lee S.G., Do H., Park J.C., Kim E., Choe Y.H., Han S.J.,
RA Kim H.J.;
RT "Optimization of the pilot-scale production of an ice-binding protein by
RT fed-batch culture of Pichia pastoris.";
RL Appl. Microbiol. Biotechnol. 97:3383-3393(2013).
RN [5]
RP CRYSTALLIZATION.
RX PubMed=21795798; DOI=10.1107/s1744309111018446;
RA Park A.K., Park K.S., Kim H.J., Park H., Ahn I.Y., Chi Y.M., Moon J.H.;
RT "Crystallization and preliminary X-ray crystallographic studies of the ice-
RT binding protein from the Arctic [correction of Aantarctic] yeast
RT Leucosporidium sp. AY30.";
RL Acta Crystallogr. F 67:800-802(2011).
RN [6]
RP FUNCTION, AND BIOTECHNOLOGY.
RX PubMed=22622645; DOI=10.1007/s12010-012-9739-z;
RA Lee S.G., Koh H.Y., Lee J.H., Kang S.H., Kim H.J.;
RT "Cryopreservative effects of the recombinant ice-binding protein from the
RT arctic yeast Leucosporidium sp. on red blood cells.";
RL Appl. Biochem. Biotechnol. 167:824-834(2012).
RN [7] {ECO:0007744|PDB:3UYU, ECO:0007744|PDB:3UYV}
RP X-RAY CRYSTALLOGRAPHY (1.57 ANGSTROMS) OF 21-261 OF NON-GLYCOSYLATED AND
RP GLYCOSYLATED PROTEINS, FUNCTION, SUBUNIT, GLYCOSYLATION AT ASN-185,
RP MUTAGENESIS OF SER-43; ALA-49; THR-65; THR-109; SER-147; SER-171; THR-198;
RP THR-216; SER-222; ALA-234; THR-239 AND 244-PHE--LEU-261, AND CIRCULAR
RP DICHROISM ANALYSIS.
RC STRAIN=AY30 {ECO:0000303|PubMed:22303017};
RX PubMed=22303017; DOI=10.1074/jbc.m111.331835;
RA Lee J.H., Park A.K., Do H., Park K.S., Moh S.H., Chi Y.M., Kim H.J.;
RT "Structural basis for antifreeze activity of ice-binding protein from
RT arctic yeast.";
RL J. Biol. Chem. 287:11460-11468(2012).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.34 ANGSTROMS) OF 21-53; 52-97 AND 100-243 OF
RP CHIMERA WITH BACTERIAL ANTIFREEZE PROTEIN, FUNCTION, SUBUNIT, AND CIRCULAR
RP DICHROISM ANALYSIS.
RX PubMed=24699650; DOI=10.1107/s1399004714000996;
RA Do H., Kim S.J., Kim H.J., Lee J.H.;
RT "Structure-based characterization and antifreeze properties of a
RT hyperactive ice-binding protein from the Antarctic bacterium Flavobacterium
RT frigoris PS1.";
RL Acta Crystallogr. D 70:1061-1073(2014).
CC -!- FUNCTION: Confers freeze tolerance. Binds to the surface of ice
CC crystals and inhibits their growth. Has low thermal hysteresis (TH)
CC activity, which is the ability to lower the freezing point of an
CC aqueous solution below its melting point (PubMed:24699650,
CC PubMed:22303017, PubMed:20067781, PubMed:22426061, PubMed:23203635,
CC PubMed:22622645). The TH activity of this protein is approximately 0.2
CC degrees Celsius at 50 uM and 0.3 degrees Celsius at 400 uM
CC (PubMed:24699650). {ECO:0000269|PubMed:20067781,
CC ECO:0000269|PubMed:22303017, ECO:0000269|PubMed:22426061,
CC ECO:0000269|PubMed:22622645, ECO:0000269|PubMed:23203635,
CC ECO:0000269|PubMed:24699650}.
CC -!- SUBUNIT: Homodimer (PubMed:22426061, PubMed:24699650, PubMed:22303017).
CC Dimerization is not required for the thermal hysteresis (TH) activity
CC (PubMed:22303017). {ECO:0000269|PubMed:22303017,
CC ECO:0000269|PubMed:22426061, ECO:0000269|PubMed:24699650}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:20067781,
CC ECO:0000269|PubMed:22426061, ECO:0000269|PubMed:23203635}.
CC -!- PTM: Glycosylated (PubMed:20067781, PubMed:22426061, PubMed:22303017,
CC PubMed:23203635). Glycosylation is not required for the thermal
CC hysteresis (TH) activity (PubMed:22426061). Glycosylation may increase
CC stability and secretion of this protein (Probable).
CC {ECO:0000269|PubMed:20067781, ECO:0000269|PubMed:22303017,
CC ECO:0000269|PubMed:22426061, ECO:0000269|PubMed:23203635,
CC ECO:0000305|PubMed:22303017}.
CC -!- MASS SPECTROMETRY: Mass=25565; Method=MALDI; Note=The measured mass is
CC that of the mature protein.; Evidence={ECO:0000269|PubMed:23203635};
CC -!- BIOTECHNOLOGY: Improves cryopreservation of the human red blood cells
CC (RBCs). Almost 90 % hemolysis damage induced to RBCs by freeze thawing
CC with the slow warming at 22 degrees Celsius is dramatically decreased
CC to less than 16 % when 0.4 or 0.8 mg/ml of this protein is used as a
CC cryoprotectant. The post-thaw cell counts of RBCs frozen in the
CC presence of 0.8 mg/ml of this protein and thawed by slow warming at 22
CC degrees Celsius is almost the same as that of non-frozen intact RBCs,
CC whereas the recovery of RBCs frozen in its absence is less than 30 %.
CC {ECO:0000269|PubMed:22622645}.
CC -!- SIMILARITY: Belongs to the ice-binding protein family. {ECO:0000305}.
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=Ice whisperer - Issue 209 of
CC December 2018;
CC URL="https://web.expasy.org/spotlight/back_issues/209/";
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DR EMBL; GQ336994; ACU30806.1; -; Genomic_DNA.
DR EMBL; GQ336995; ACU30807.1; -; mRNA.
DR PDB; 3UYU; X-ray; 1.57 A; A/B=21-261.
DR PDB; 3UYV; X-ray; 2.43 A; A=21-261.
DR PDB; 4NU3; X-ray; 1.40 A; A/B=52-97.
DR PDB; 4NUH; X-ray; 1.34 A; A=21-53, A=100-243.
DR PDBsum; 3UYU; -.
DR PDBsum; 3UYV; -.
DR PDBsum; 4NU3; -.
DR PDBsum; 4NUH; -.
DR AlphaFoldDB; C7F6X3; -.
DR SMR; C7F6X3; -.
DR iPTMnet; C7F6X3; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0050825; F:ice binding; IDA:UniProtKB.
DR InterPro; IPR021884; Ice-bd_prot.
DR Pfam; PF11999; Ice_binding; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antifreeze protein; Direct protein sequencing; Glycoprotein;
KW Secreted; Signal; Stress response.
FT SIGNAL 1..20
FT /evidence="ECO:0000255, ECO:0000269|PubMed:22426061,
FT ECO:0000269|PubMed:23203635"
FT CHAIN 21..261
FT /note="Ice-binding protein"
FT /evidence="ECO:0000255"
FT /id="PRO_5011088933"
FT SITE 65
FT /note="Ice-binding"
FT /evidence="ECO:0000305|PubMed:22303017"
FT SITE 147
FT /note="Ice-binding"
FT /evidence="ECO:0000305|PubMed:22303017"
FT SITE 234
FT /note="Ice-binding"
FT /evidence="ECO:0000305|PubMed:22303017"
FT CARBOHYD 185
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:22303017,
FT ECO:0007744|PDB:3UYV"
FT MUTAGEN 43
FT /note="S->Y: Has 35% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 49
FT /note="A->Y: Has 89% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 65
FT /note="T->Y: Has 28% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 109
FT /note="T->Y: Has 92% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 147
FT /note="S->Y: Has 62% thermal hysteresis (TH) activity of
FT that of the wild-type. Loss of TH activity; when associated
FT with Y-234."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 171
FT /note="S->Y: Has 67% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 198
FT /note="T->Y: Has 50% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 216
FT /note="T->Y: Has 59% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 222
FT /note="S->Y: Has 64% thermal hysteresis (TH) activity of
FT that of the wild-type."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 234
FT /note="A->Y: Has 47% thermal hysteresis (TH) activity of
FT that of the wild-type. Loss of TH activity; when associated
FT with Y-147."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 239
FT /note="T->Y: No effect on thermal hysteresis (TH)
FT activity."
FT /evidence="ECO:0000269|PubMed:22303017"
FT MUTAGEN 244..261
FT /note="Missing: Has 13% higher thermal hysteresis (TH)
FT activity compared to the wild-type. Loss of
FT homodimerization."
FT /evidence="ECO:0000269|PubMed:22303017"
FT HELIX 29..33
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 35..46
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 49..52
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 54..59
FT /evidence="ECO:0007829|PDB:3UYU"
FT HELIX 61..63
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 64..67
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 83..85
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:3UYU"
FT HELIX 96..116
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 121..124
FT /evidence="ECO:0007829|PDB:3UYU"
FT HELIX 125..128
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 132..135
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 137..144
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 146..148
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 152..155
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 162..168
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 170..172
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 177..181
FT /evidence="ECO:0007829|PDB:3UYU"
FT HELIX 186..188
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 189..195
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 197..199
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 204..213
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 215..217
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 222..237
FT /evidence="ECO:0007829|PDB:3UYU"
FT STRAND 239..241
FT /evidence="ECO:0007829|PDB:3UYU"
SQ SEQUENCE 261 AA; 26807 MW; F114BC33CBC9B469 CRC64;
MSLLSIITIG LAGLGGLVNG QRDLSVELGV ASNFAILAKA GISSVPDSAI LGDIGVSPAA
ATYITGFGLT QDSSTTYATS PQVTGLIYAA DYSTPTPNYL AAAVANAETA YNQAAGFVDP
DFLELGAGEL RDQTLVPGLY KWTSSVSVPT DLTFEGNGDA TWVFQIAGGL SLADGVAFTL
AGGANSTNIA FQVGDDVTVG KGAHFEGVLL AKRFVTLQTG SSLNGRVLSQ TEVALQKATV
NSPFVPAPEV VQKRSNARQW L
