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 07-NOV-2018, entry version 30. 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. 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. RT AY30."; RL Submitted (JUN-2009) to the EMBL/GenBank/DDBJ databases. RN [2] RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 214-226, RP IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, SUBCELLULAR LOCATION, RP 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 RT psychrophilic 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, RP GLYCOSYLATION, 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 RT by 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., RA Moon J.H.; RT "Crystallization and preliminary X-ray crystallographic studies of the RT ice-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 RT the arctic yeast Leucosporidium sp. on red blood cells."; RL Appl. Biochem. Biotechnol. 167:824-834(2012). RN [7] {ECO:0000244|PDB:3UYU, ECO:0000244|PDB:3UYV} RP X-RAY CRYSTALLOGRAPHY (1.57 ANGSTROMS) OF 21-261 OF NON-GLYCOSYLATED RP AND GLYCOSYLATED PROTEINS, FUNCTION, SUBUNIT, GLYCOSYLATION AT RP ASN-185, MUTAGENESIS OF SER-43; ALA-49; THR-65; THR-109; SER-147; RP SER-171; THR-198; THR-216; SER-222; ALA-234; THR-239 AND RP 244-PHE--LEU-261, AND CIRCULAR 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 RP CIRCULAR 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 RT Flavobacterium 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 CC (TH) activity, which is the ability to lower the freezing point of CC an aqueous solution below its melting point (PubMed:24699650, CC PubMed:22303017, PubMed:20067781, PubMed:22426061, CC PubMed:23203635, PubMed:22622645). The TH activity of this protein CC is approximately 0.2 degrees Celsius at 50 uM and 0.3 degrees CC Celsius at 400 uM (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, CC PubMed:22303017). Dimerization is not required for the thermal CC hysteresis (TH) activity (PubMed:22303017). CC {ECO:0000269|PubMed:22303017, ECO:0000269|PubMed:22426061, CC 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, CC PubMed:22303017, PubMed:23203635). Glycosylation is not required CC for the thermal hysteresis (TH) activity (PubMed:22426061). CC Glycosylation may increase stability and secretion of this protein CC (Probable). {ECO:0000269|PubMed:20067781, CC ECO:0000269|PubMed:22303017, ECO:0000269|PubMed:22426061, CC ECO:0000269|PubMed:23203635, ECO:0000305|PubMed:22303017}. CC -!- MASS SPECTROMETRY: Mass=25565; Method=MALDI; Range=20-261; CC Note=The measured mass is that of the mature protein.; CC Evidence={ECO:0000269|PubMed:23203635}; CC -!- BIOTECHNOLOGY: Improves cryopreservation of the human red blood CC cells (RBCs). Almost 90 % hemolysis damage induced to RBCs by CC freeze thawing with the slow warming at 22 degrees Celsius is CC dramatically decreased to less than 16 % when 0.4 or 0.8 mg/ml of CC this protein is used as a cryoprotectant. The post-thaw cell CC counts of RBCs frozen in the presence of 0.8 mg/ml of this protein CC and thawed by slow warming at 22 degrees Celsius is almost the CC same as that of non-frozen intact RBCs, whereas the recovery of CC RBCs frozen in its absence is less than 30 %. CC {ECO:0000269|PubMed:22622645}. CC -!- SIMILARITY: Belongs to the ice-binding protein family. CC {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- 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 SMR; 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-like. DR Pfam; PF11999; DUF3494; 1. PE 1: Evidence at protein level; KW 3D-structure; Antifreeze protein; Direct protein sequencing; KW Glycoprotein; Secreted; Signal; Stress response. FT SIGNAL 1 20 {ECO:0000255, FT ECO:0000269|PubMed:22426061, FT ECO:0000269|PubMed:23203635}. FT CHAIN 21 261 Ice-binding protein. {ECO:0000255}. FT /FTId=PRO_5011088933. FT SITE 65 65 Ice-binding. FT {ECO:0000305|PubMed:22303017}. FT SITE 147 147 Ice-binding. FT {ECO:0000305|PubMed:22303017}. FT SITE 234 234 Ice-binding. FT {ECO:0000305|PubMed:22303017}. FT CARBOHYD 185 185 N-linked (GlcNAc...) asparagine. FT {ECO:0000244|PDB:3UYV, FT ECO:0000269|PubMed:22303017}. FT MUTAGEN 43 43 S->Y: Has 35% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 49 49 A->Y: Has 89% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 65 65 T->Y: Has 28% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 109 109 T->Y: Has 92% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 147 147 S->Y: Has 62% thermal hysteresis (TH) FT activity of that of the wild-type. Loss FT of TH activity; when associated with Y- FT 234. {ECO:0000269|PubMed:22303017}. FT MUTAGEN 171 171 S->Y: Has 67% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 198 198 T->Y: Has 50% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 216 216 T->Y: Has 59% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 222 222 S->Y: Has 64% thermal hysteresis (TH) FT activity of that of the wild-type. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 234 234 A->Y: Has 47% thermal hysteresis (TH) FT activity of that of the wild-type. Loss FT of TH activity; when associated with Y- FT 147. {ECO:0000269|PubMed:22303017}. FT MUTAGEN 239 239 T->Y: No effect on thermal hysteresis FT (TH) activity. FT {ECO:0000269|PubMed:22303017}. FT MUTAGEN 244 261 Missing: Has 13% higher thermal FT hysteresis (TH) activity compared to the FT wild-type. Loss of homodimerization. FT {ECO:0000269|PubMed:22303017}. FT HELIX 29 33 {ECO:0000244|PDB:3UYU}. FT STRAND 35 46 {ECO:0000244|PDB:3UYU}. FT STRAND 49 52 {ECO:0000244|PDB:3UYU}. FT STRAND 54 59 {ECO:0000244|PDB:3UYU}. FT HELIX 61 63 {ECO:0000244|PDB:3UYU}. FT STRAND 64 67 {ECO:0000244|PDB:3UYU}. FT STRAND 83 85 {ECO:0000244|PDB:3UYU}. FT STRAND 87 89 {ECO:0000244|PDB:3UYU}. FT HELIX 96 116 {ECO:0000244|PDB:3UYU}. FT STRAND 121 124 {ECO:0000244|PDB:3UYU}. FT HELIX 125 128 {ECO:0000244|PDB:3UYU}. FT STRAND 132 135 {ECO:0000244|PDB:3UYU}. FT STRAND 137 144 {ECO:0000244|PDB:3UYU}. FT STRAND 146 148 {ECO:0000244|PDB:3UYU}. FT STRAND 152 155 {ECO:0000244|PDB:3UYU}. FT STRAND 162 168 {ECO:0000244|PDB:3UYU}. FT STRAND 170 172 {ECO:0000244|PDB:3UYU}. FT STRAND 177 181 {ECO:0000244|PDB:3UYU}. FT HELIX 186 188 {ECO:0000244|PDB:3UYU}. FT STRAND 189 195 {ECO:0000244|PDB:3UYU}. FT STRAND 197 199 {ECO:0000244|PDB:3UYU}. FT STRAND 204 213 {ECO:0000244|PDB:3UYU}. FT STRAND 215 217 {ECO:0000244|PDB:3UYU}. FT STRAND 222 237 {ECO:0000244|PDB:3UYU}. FT STRAND 239 241 {ECO:0000244|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 //