1z68: Difference between revisions

Revision as of 17:12, 21 February 2008

Crystal Structure Of Human Fibroblast Activation Protein alpha

OverviewOverview

Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha.

About this StructureAbout this Structure

1Z68 is a Single protein structure of sequence from Homo sapiens with and as ligands. Full crystallographic information is available from OCA.

ReferenceReference

Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha., Aertgeerts K, Levin I, Shi L, Snell GP, Jennings A, Prasad GS, Zhang Y, Kraus ML, Salakian S, Sridhar V, Wijnands R, Tennant MG, J Biol Chem. 2005 May 20;280(20):19441-4. Epub 2005 Apr 4. PMID:15809306

Page seeded by OCA on Thu Feb 21 16:12:26 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 1: / Line 1: @@
-[[Image:1z68.gif|left|200px]]<br />
+[[Image:1z68.gif|left|200px]]<br /><applet load="1z68" size="350" color="white" frame="true" align="right" spinBox="true"
-<applet load="1z68" size="450" color="white" frame="true" align="right" spinBox="true"
 caption="1z68, resolution 2.60&Aring;" />
 '''Crystal Structure Of Human Fibroblast Activation Protein alpha'''<br />
 ==Overview==
-Fibroblast activation protein alpha (FAPalpha) is highly expressed in, epithelial cancers and has been implicated in extracellular matrix, remodeling, tumor growth, and metastasis. We present the first high, resolution structure for the apoenzyme as well as kinetic data toward, small dipeptide substrates. FAPalpha exhibits a dipeptidyl peptidase IV, (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an, eight-bladed beta-propeller domain. Known DPPIV dipeptides are cleaved by, FAPalpha with an approximately 100-fold decrease in catalytic efficiency, compared with DPPIV. Moreover, FAPalpha, but not DPPIV, possesses, endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked, peptides. Comparison of the crystal structures of FAPalpha and DPPIV, revealed one major difference in the vicinity of the Glu motif, (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the, active site of the enzyme. Ala(657) in FAPalpha, instead of Asp(663) as in, DP-PIV, reduces the acidity in this pocket, and this change could explain, the lower affinity for N-terminal amines by FAPalpha. This hypothesis was, tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on, average an approximately 60-fold increase in the catalytic efficiency, as, measured by k(cat)/K(m), for the cleavage of dipeptide substrates., Furthermore, the catalytic efficiency of the mutant is reduced by, approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin., Our data provide a clear understanding of the molecular determinants, responsible for the substrate specificity and endopeptidase activity of, FAPalpha.
+Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha.
 ==About this Structure==
-Z68 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 NAG and NDG as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1Z68 OCA].
+Z68 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=NAG:'>NAG</scene> and <scene name='pdbligand=NDG:'>NDG</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Z68 OCA].
 ==Reference==
@@ Line 15: / Line 14: @@
 [[Category: Single protein]]
 [[Category: Aertgeerts, K.]]
-[[Category: Kraus, M.L.]]
+[[Category: Kraus, M L.]]
 [[Category: Levin, I.]]
-[[Category: Prasad, G.S.]]
+[[Category: Prasad, G S.]]
 [[Category: Salakian, S.]]
 [[Category: Shi, L.]]
-[[Category: Snell, G.P.]]
+[[Category: Snell, G P.]]
 [[Category: Sridhar, V.]]
-[[Category: Tennant, M.G.]]
+[[Category: Tennant, M G.]]
 [[Category: Wijnands, R.]]
 [[Category: Zhang, Y.]]
@@ Line 34: / Line 33: @@
 [[Category: seprase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 20:29:49 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:12:26 2008''