title: Molecular basis of LMAN1 in coordinating LMAN1-MCFD2 cargo receptor formation and ER-to-Golgi transport of FV/FVIII.
abstract: The LMAN1-MCFD2 (lectin, mannose binding 1/multiple coagulation factor deficiency protein 2) cargo receptor complex transports coagulation factors V (FV) and VIII (FVIII) from the endoplasmic reticulum (ER) to the ER-Golgi intermediate compartment (ERGIC). LMAN1 (ERGIC-53) is a hexameric transmembrane protein with a carbohydrate recognition domain (CRD) on the ER luminal side. Here, we show that mutations in the first beta sheet of the CRD abolish MCFD2 binding without affecting the mannose binding, suggesting that LMAN1 interacts with MCFD2 through its N-terminal beta sheet, consistent with recently reported crystal structures of the CRD-MCFD2 complex. Mutations in the Ca(2+)- and sugar-binding sites of the CRD disrupt FV and FVIII interactions, without affecting MCFD2 binding. This interaction is independent of MCFD2, as LMAN1 mutants defective in MCFD2 binding can still interact with FVIII. Thus, the CRD of LMAN1 contains distinct, separable binding sites for both its partner protein (MCFD2) and the cargo proteins (FV/FVIII). Monomeric LMAN1 mutants are defective in ER exit and unable to interact with MCFD2, suggesting that the oligomerization of LMAN1 is necessary for its cargo receptor function. These results point to a central role of LMAN1 in regulating the binding in the ER and the subsequent release in the ERGIC of FV and FVIII.
authors.affiliation: Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Zheng
authors.forename: Hui-Hui
authors.initials: HH
authors.lastname: Liu
authors.forename: Shuguang
authors.initials: S
authors.lastname: Yuan
authors.forename: Jiahai
authors.initials: J
authors.lastname: Zhou
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D001692:Biological Transport; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005165:Factor V / Q000378:metabolism*; D005169:Factor VIII / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D037241:Mannose-Binding Lectins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064067:Rho Guanine Nucleotide Exchange Factors; D017386:Sequence Homology, Amino Acid
chemical_list: D020662:Guanine Nucleotide Exchange Factors; C084004:LMAN1 protein, human; C577400:MCF2L protein, human; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D012333:RNA, Messenger; D064067:Rho Guanine Nucleotide Exchange Factors; D005165:Factor V; D005169:Factor VIII; D002118:Calcium
references.citation: Cell. 1998 Apr 3;93(1):61-70
references.pmid: 9546392
references.citation: J Biol Chem. 2004 Mar 26;279(13):12943-50
references.pmid: 14718532
references.citation: Nat Cell Biol. 2010 Jan;12(1):41-6; sup pp 1-8
references.pmid: 19966784
references.citation: Eur J Biochem. 1999 Mar;260(2):392-7
references.pmid: 10095773
references.citation: Blood. 2008 Feb 15;111(4):1972-9
references.pmid: 18056485
references.citation: Mol Biol Cell. 1997 Aug;8(8):1501-12
references.pmid: 9285821
references.citation: Mol Biol Cell. 1996 Mar;7(3):483-93
references.pmid: 8868475
references.citation: Cell. 2003 Aug 22;114(4):483-95
references.pmid: 12941276
references.citation: Blood. 2006 Mar 1;107(5):1903-7
references.pmid: 16304051
references.citation: J Cell Sci. 2003 Nov 1;116(Pt 21):4429-40
references.pmid: 13130098
references.citation: Haemophilia. 2007 Jul;13(4):413-9
references.pmid: 17610559
references.citation: J Biol Chem. 2005 Jul 8;280(27):25881-6
references.pmid: 15886209
references.citation: Nat Cell Biol. 1999 Oct;1(6):330-4
references.pmid: 10559958
references.citation: FEBS Lett. 2010 Mar 5;584(5):878-82
references.pmid: 20138881
references.citation: Cell. 2003 Aug 22;114(4):497-509
references.pmid: 12941277
references.citation: J Mol Biol. 2005 Dec 2;354(3):556-68
references.pmid: 16257008
references.citation: FEBS Lett. 2009 Dec 3;583(23):3796-803
references.pmid: 19850039
references.citation: Mol Biol Cell. 2003 Jul;14(7):3055-63
references.pmid: 12857885
references.citation: J Cell Biol. 2008 Feb 25;180(4):705-12
references.pmid: 18283111
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4034-9
references.pmid: 20142513
references.citation: Biotechniques. 2006 Jan;40(1):61-6
references.pmid: 16454041
references.citation: J Biol Chem. 1999 Nov 12;274(46):32539-42
references.pmid: 10551804
references.citation: Blood. 2010 Feb 4;115(5):1081-7
references.pmid: 20007547
references.citation: Nat Genet. 2003 Jun;34(2):220-5
references.pmid: 12717434
references.citation: J Biol Chem. 2005 Mar 4;280(9):7758-68
references.pmid: 15623526
references.citation: J Thromb Haemost. 2004 Sep;2(9):1564-72
references.pmid: 15333032
references.citation: Trends Biochem Sci. 2007 Aug;32(8):381-8
references.pmid: 17618120
references.citation: J Biol Chem. 2001 Aug 31;276(35):33027-35
references.pmid: 11402049
references.citation: Br J Haematol. 2009 Apr;145(1):15-23
references.pmid: 19183188
references.citation: Am J Hematol. 2009 Nov;84(11):738-42
references.pmid: 19787799
references.citation: Annu Rev Cell Dev Biol. 2004;20:87-123
references.pmid: 15473836
references.citation: J Mol Biol. 2003 Dec 12;334(5):845-51
references.pmid: 14643651
references.citation: Proc Natl Acad Sci U S A. 2005 May 3;102(18):6350-5
references.pmid: 15849265
references.citation: J Biol Chem. 2002 May 3;277(18):15979-84
references.pmid: 11850423
references.citation: Blood. 2008 Jun 15;111(12):5592-600
references.pmid: 18391077
references.citation: J Cell Biol. 1998 Jul 27;142(2):377-89
references.pmid: 9679138
references.citation: J Cell Sci. 2000 Feb;113 ( Pt 4):587-96
references.pmid: 10652252
references.citation: Traffic. 2006 Nov;7(11):1473-81
references.pmid: 17010120
references.citation: Mol Cell. 2002 Apr;9(4):789-98
references.pmid: 11983170
references.citation: J Mol Biol. 2008 Sep 12;381(4):941-55
references.pmid: 18590741