US 12,012,519 B2
Liquid-liquid phase separation driven protein-based underwater adhesive coatings
Chao Zhong, Shanghai (CN); and Mengkui Cui, Shanghai (CN)
Assigned to ShanghaiTech University, Shanghai (CN)
Filed by ShanghaiTech University, Shanghai (CN)
Filed on Feb. 8, 2021, as Appl. No. 17/169,581.
Application 17/169,581 is a continuation of application No. PCT/CN2019/100159, filed on Aug. 12, 2019.
Prior Publication US 2021/0155826 A1, May 27, 2021
Int. Cl. C12N 15/62 (2006.01); B82Y 30/00 (2011.01); C07K 14/435 (2006.01); C09D 189/00 (2006.01)
CPC C09D 189/00 (2013.01) [C07K 14/435 (2013.01); C07K 2319/20 (2013.01)] 20 Claims
 
1. A dimeric protein comprising a marine adhesive protein (MAP) domain and a liquid-liquid phase separation (LLPS) mediating low complexity (LC) domain;
wherein the marine adhesive protein (MAP) is a polypeptide selected from the group consisting of:
a) a mussel foot protein selected from the group consisting of: mfp5, mfp3 and mfp3s;
b) a barnacle adhesive protein selected from the group consisting of: cp19k, cp20k, and cp68k; and
c) a sandcastle worm adhesive protein selected from the group consisting of: Pc2 and Pc3;
wherein the low complexity (LC) domain is a polypeptide selected from the group consisting of:
LC domain TAR DNA-binding protein 43 (TDP-43) with amino acid sequence of
(SEQ ID NO: 01)
KHNSNRQLERSGREGGNPGGFGNQGGFGNSRGGGAGLGNNQGSNMGGGM
 
NFGAFSINPAMMAAAQAALQSSWGMMGMLASQQNQSGPSGNNQNQGNMQ
 
REPNQAFGSGNNSYSGSNSGAAIGWGSASNAGSGSGFNGGFGSSMDSKS
 
SGWGM;
heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) with amino acid sequence of
(SEQ ID NO: 03)
MASASSSQRGRSGSGNFGGGRGGGFGGNDNFGRGGNFSGRGGFGGSRGG
 
GGYGGSGDGYNGFGNDGSNFGGGGSYNDFGNYNNQSSNFGPMKGGNFGG
 
RSSGPYGGGGQYFAKPRNQGGYGGSSSSSSYGSGRRF;
heterogeneous nuclear ribonucleoprotein A2 (hnRNPA2) with amino acid sequence of
(SEQ ID NO: 04)
MQEVQSSRSGRGGNFGFGDSRGGGGNFGPGPGSNFRGGSDGYGSGRGFG
 
DGYNGYGGGPGGGNFGGSPGYGGGRGGYGGGGPGYGNQGGGYGGGYDNY
 
GGGNYGSGNYNDFGNYNQQPSNYGPMKSGNFGGSRNMGGPYGGGNYGPG
 
GSGGSGGYGGRSRY;
cytotoxic granule-associated RNA binding protein TIA1 with amino acid sequence of
(SEQ ID NO: 05)
INPVQQQNQIGYPQPYGQWGQWYGNAQQIGQYMPNGWQVPAYGMYGQAW
 
NQQGENQTQSSAPWMGPNYGVQPPQGQNGSMLPNQPSGYRVAGYETQ;
cytoplasmic polyadenylation element binding protein 2 (CPEB2) with amino acid sequence of
(SEQ ID NO: 06)
PPPSPDSENGFYPGLPSSMNPAFFPSFSPVSPHGCTGLSVPTSGGGGGG
 
FGGPFSATAVPPPPPPAMNIPQQQPPPPAAPQQPQSRRSPVSPQLQQQH
 
QAAAAAFLQQRNSYNHHQPLLKQSPWSNHQSSGWGTGSM;
fragile X mental retardation protein (FMRP) with amino acid sequence of
(SEQ ID NO: 07)
GQGMGRGSRPYRNRGHGRRGPGYTSGTNSEASNASETESDHRDELSDWS
 
LAPTEEERESFLRRGDGRRRGGGGRGQGGRGRGGGFKGNDDHSRTDNRP
 
RNPREAKGRTTDGSLQIRVDCNNERSVHTKTLQNTSSEGSRLRTGKDRN
 
QKKEKPDSVDGQQPLVNGVP;
cold inducible RNA binding protein (CIRBP) with amino acid sequence of
(SEQ ID NO: 08)
MASDEGKLFVGGLSFDTNEQSLEQVFSKYGQISEVVVVKDRETQRSRGF
 
GFVTFENIDDAKDAMMAMNGKSVDGRQIRVDQAGKSSDNRSRGYRGGSA
 
GGRGFFRGGRGRGRGFSRGGGDRGYGGNRFESRSGGYGGSRDYYSSRSQ
 
SGGYSDRSSGGSYRDSYDSYATHNE;
RNA binding motif protein 3 (RBM3) with amino acid sequence of
(SEQ ID NO: 09)
MSSEEGKLFVGGLNFNTDEQALEDHFSSFGPISEVVVVKDRETQRSRGF
 
GFITFTNPEHASVAMRAMNGESLDGRQIRVDHAGKSARGTRGGGFGAHG
 
RGRSYSRGGGDQGYGSGRYYDSRPGGYGYGYGRSRDYNGRNQGGYDRYS
 
GGNYRDNYDN;
and
yeast Sup35 with amino acid sequence of
(SEQ ID NO: 10)
SDSNQGNNQQNYQQYSQNGNQQQGNNRYQGYQAYNAQAQPAGGYYQNYQ
 
GYSGYQQGGYQQYNPDAGYQQQYNPQGGYQQYNPQGGYQQQFNPQGGRG
 
NYKNFNYNNSLQGYQAGFQPQSQGMSLNDFQKQQKQ.