1ffm: Difference between revisions

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[[Image:1ffm.gif|left|200px]]<br />

[[Image:1ffm.gif|left|200px]]<br /><applet load="1ffm" size="350" color="white" frame="true" align="right" spinBox="true"

<applet load="1ffm" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

caption="1ffm" />

'''THE FIRST EGF-LIKE DOMAIN FROM HUMAN BLOOD COAGULATION FVII (FUCOSYLATED AT SER-60), NMR, MINIMIZED AVERAGE STRUCTURE'''<br />

==Overview==

The first epidermal growth factor-like domain (EGF-1) from blood, coagulation factor VII (FVII) contains two unusual O-linked glycosylation, sites at Ser-52 and Ser-60. We report here a detailed study of the effect, of O-fucosylation at Ser-60 on the structure of FVII EGF-1, its, Ca2+-binding affinity, and its interaction with tissue factor (TF). The in, vitro fucosylation of the nonglycosylated FVII EGF-1 was achieved by using, O-fucosyltransferase purified from Chinese hamster ovary cells. Distance, and dihedral constraints derived from NMR data were used to determine the, solution structures of both nonglycosylated and fucosylated FVII EGF-1 in, the presence of CaCl2. The overall structure of fucosylated FVII EGF-1 is, very similar to the nonfucosylated form even for the residues near the, fucosylation site. The Ca2+ dissociation constants (Kd) for the, nonfucosylated and fucosylated FVII EGF-1 were found to be 16.4 +/- 1.8, and 8.6 +/- 1.4 mM, respectively. The FVII EGF-1 domain binds to the, extracellular part of TF with a low affinity (Kd approximately 0. 6 mM), and the addition of fucose appears to have no effect on this affinity., These results indicate that the FVII EGF-1 alone cannot form a tight, complex with TF and suggest that the high binding affinity of FVIIa for TF, requires cooperative interaction among the four domains in FVII with TF., Although the fucose has no significant effect on the interaction between, TF and the individual FVII EGF-1 domain, it may affect the interaction of, full-length FVIIa with TF by influencing its Ca2+-binding affinity.

The first epidermal growth factor-like domain (EGF-1) from blood coagulation factor VII (FVII) contains two unusual O-linked glycosylation sites at Ser-52 and Ser-60. We report here a detailed study of the effect of O-fucosylation at Ser-60 on the structure of FVII EGF-1, its Ca2+-binding affinity, and its interaction with tissue factor (TF). The in vitro fucosylation of the nonglycosylated FVII EGF-1 was achieved by using O-fucosyltransferase purified from Chinese hamster ovary cells. Distance and dihedral constraints derived from NMR data were used to determine the solution structures of both nonglycosylated and fucosylated FVII EGF-1 in the presence of CaCl2. The overall structure of fucosylated FVII EGF-1 is very similar to the nonfucosylated form even for the residues near the fucosylation site. The Ca2+ dissociation constants (Kd) for the nonfucosylated and fucosylated FVII EGF-1 were found to be 16.4 +/- 1.8 and 8.6 +/- 1.4 mM, respectively. The FVII EGF-1 domain binds to the extracellular part of TF with a low affinity (Kd approximately 0. 6 mM), and the addition of fucose appears to have no effect on this affinity. These results indicate that the FVII EGF-1 alone cannot form a tight complex with TF and suggest that the high binding affinity of FVIIa for TF requires cooperative interaction among the four domains in FVII with TF. Although the fucose has no significant effect on the interaction between TF and the individual FVII EGF-1 domain, it may affect the interaction of full-length FVIIa with TF by influencing its Ca2+-binding affinity.

==Disease==

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==About this Structure==

1FFM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with FUC as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=1FFM OCA].

1FFM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=FUC:'>FUC</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1FFM OCA].

==Reference==

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[[Category: Homo sapiens]]

[[Category: Single protein]]

[[Category: Kao, Y.H.]]

[[Category: Kao, Y H.]]

[[Category: Kelley, R.F.]]

[[Category: Kelley, R F.]]

[[Category: Lee, G.F.]]

[[Category: Lee, G F.]]

[[Category: Lerner, L.]]

[[Category: Spellman, M.W.]]

[[Category: Spellman, M W.]]

[[Category: Starovasnik, M.A.]]

[[Category: Starovasnik, M A.]]

[[Category: Wang, Y.]]

[[Category: FUC]]

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[[Category: o- linked fucose]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Mon Nov~~ 12 16:52:~~41 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:38:08 2008''

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-[[Image:1ffm.gif|left|200px]]<br />
+[[Image:1ffm.gif|left|200px]]<br /><applet load="1ffm" size="350" color="white" frame="true" align="right" spinBox="true"
-<applet load="1ffm" size="450" color="white" frame="true" align="right" spinBox="true"
 caption="1ffm" />
 '''THE FIRST EGF-LIKE DOMAIN FROM HUMAN BLOOD COAGULATION FVII (FUCOSYLATED AT SER-60), NMR, MINIMIZED AVERAGE STRUCTURE'''<br />
 ==Overview==
-The first epidermal growth factor-like domain (EGF-1) from blood, coagulation factor VII (FVII) contains two unusual O-linked glycosylation, sites at Ser-52 and Ser-60. We report here a detailed study of the effect, of O-fucosylation at Ser-60 on the structure of FVII EGF-1, its, Ca2+-binding affinity, and its interaction with tissue factor (TF). The in, vitro fucosylation of the nonglycosylated FVII EGF-1 was achieved by using, O-fucosyltransferase purified from Chinese hamster ovary cells. Distance, and dihedral constraints derived from NMR data were used to determine the, solution structures of both nonglycosylated and fucosylated FVII EGF-1 in, the presence of CaCl2. The overall structure of fucosylated FVII EGF-1 is, very similar to the nonfucosylated form even for the residues near the, fucosylation site. The Ca2+ dissociation constants (Kd) for the, nonfucosylated and fucosylated FVII EGF-1 were found to be 16.4 +/- 1.8, and 8.6 +/- 1.4 mM, respectively. The FVII EGF-1 domain binds to the, extracellular part of TF with a low affinity (Kd approximately 0. 6 mM), and the addition of fucose appears to have no effect on this affinity., These results indicate that the FVII EGF-1 alone cannot form a tight, complex with TF and suggest that the high binding affinity of FVIIa for TF, requires cooperative interaction among the four domains in FVII with TF., Although the fucose has no significant effect on the interaction between, TF and the individual FVII EGF-1 domain, it may affect the interaction of, full-length FVIIa with TF by influencing its Ca2+-binding affinity.
+The first epidermal growth factor-like domain (EGF-1) from blood coagulation factor VII (FVII) contains two unusual O-linked glycosylation sites at Ser-52 and Ser-60. We report here a detailed study of the effect of O-fucosylation at Ser-60 on the structure of FVII EGF-1, its Ca2+-binding affinity, and its interaction with tissue factor (TF). The in vitro fucosylation of the nonglycosylated FVII EGF-1 was achieved by using O-fucosyltransferase purified from Chinese hamster ovary cells. Distance and dihedral constraints derived from NMR data were used to determine the solution structures of both nonglycosylated and fucosylated FVII EGF-1 in the presence of CaCl2. The overall structure of fucosylated FVII EGF-1 is very similar to the nonfucosylated form even for the residues near the fucosylation site. The Ca2+ dissociation constants (Kd) for the nonfucosylated and fucosylated FVII EGF-1 were found to be 16.4 +/- 1.8 and 8.6 +/- 1.4 mM, respectively. The FVII EGF-1 domain binds to the extracellular part of TF with a low affinity (Kd approximately 0. 6 mM), and the addition of fucose appears to have no effect on this affinity. These results indicate that the FVII EGF-1 alone cannot form a tight complex with TF and suggest that the high binding affinity of FVIIa for TF requires cooperative interaction among the four domains in FVII with TF. Although the fucose has no significant effect on the interaction between TF and the individual FVII EGF-1 domain, it may affect the interaction of full-length FVIIa with TF by influencing its Ca2+-binding affinity.
 ==Disease==
@@ Line 11: / Line 10: @@
 ==About this Structure==
-FFM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with FUC as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1FFM OCA].
+FFM is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=FUC:'>FUC</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1FFM OCA].
 ==Reference==
@@ Line 17: / Line 16: @@
 [[Category: Homo sapiens]]
 [[Category: Single protein]]
-[[Category: Kao, Y.H.]]
+[[Category: Kao, Y H.]]
-[[Category: Kelley, R.F.]]
+[[Category: Kelley, R F.]]
-[[Category: Lee, G.F.]]
+[[Category: Lee, G F.]]
 [[Category: Lerner, L.]]
-[[Category: Spellman, M.W.]]
+[[Category: Spellman, M W.]]
-[[Category: Starovasnik, M.A.]]
+[[Category: Starovasnik, M A.]]
 [[Category: Wang, Y.]]
 [[Category: FUC]]
@@ Line 32: / Line 31: @@
 [[Category: o- linked fucose]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 16:52:41 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:38:08 2008''