2sfa: Difference between revisions

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[[Image:2sfa.jpg|left|200px]]<br /><applet load="2sfa" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:2sfa.jpg|left|200px]]<br /><applet load="2sfa" size="350" color="white" frame="true" align="right" spinBox="true"

caption="2sfa, resolution 1.6Å" />

'''SERINE PROTEINASE FROM STREPTOMYCES FRADIAE ATCC 14544'''<br />

==Overview==

A proteinase having wide substrate specificity was isolated from, Streptomyces fradiae ATCC 14544. This proteinase, which we propose to call, SFase-2, was purified from the culture filtrate by S-Sepharose, chromatography. The purified enzyme showed an apparent molecular mass of, 19 kDa on SDS/PAGE. When synthetic peptides were used as substrates, SFase-2 showed broad substrate specificity. It also hydrolyzed keratin, elastin and collagen as proteinaceous substrates. It was completely, inhibited by diisopropylfluorophosphate and chymostatin, but not by, tosylphenylalaninechloromethane, tosyllysinechloromethane or EDTA, indicating that it can be classified as a serine proteinase. The matured, protein sequence of SFase-2 was determined by a combination of amino acid, sequencing and the DNA sequencing of the gene. SFase-2, consisting of 191, amino acids, is a novel proteinase. It showed 76% similarity in the amino, acid sequence with Streptomyces griseus proteinase A [Johnson P. and, Smillie L. B. (1974) FEBS Lett. 47, 1-6]. For insight into the, three-dimensional structure of SFase-2, we obtained single crystals by the, vapor diffusion method using sodium phosphate as a precipitant. These, crystals belonged to the orthorhombic, space group P2(1)2(1)2(1) with cell, dimensions a = 6.92 nm, b = 7.28 nm, c = 2.99 nm; one molecule was present, in the asymmetric unit.

A proteinase having wide substrate specificity was isolated from Streptomyces fradiae ATCC 14544. This proteinase, which we propose to call SFase-2, was purified from the culture filtrate by S-Sepharose chromatography. The purified enzyme showed an apparent molecular mass of 19 kDa on SDS/PAGE. When synthetic peptides were used as substrates, SFase-2 showed broad substrate specificity. It also hydrolyzed keratin, elastin and collagen as proteinaceous substrates. It was completely inhibited by diisopropylfluorophosphate and chymostatin, but not by tosylphenylalaninechloromethane, tosyllysinechloromethane or EDTA, indicating that it can be classified as a serine proteinase. The matured protein sequence of SFase-2 was determined by a combination of amino acid sequencing and the DNA sequencing of the gene. SFase-2, consisting of 191 amino acids, is a novel proteinase. It showed 76% similarity in the amino acid sequence with Streptomyces griseus proteinase A [Johnson P. and Smillie L. B. (1974) FEBS Lett. 47, 1-6]. For insight into the three-dimensional structure of SFase-2, we obtained single crystals by the vapor diffusion method using sodium phosphate as a precipitant. These crystals belonged to the orthorhombic, space group P2(1)2(1)2(1) with cell dimensions a = 6.92 nm, b = 7.28 nm, c = 2.99 nm; one molecule was present in the asymmetric unit.

==About this Structure==

2SFA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_fradiae Streptomyces fradiae]. Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=2SFA OCA].

2SFA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_fradiae Streptomyces fradiae]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2SFA OCA].

==Reference==

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[[Category: serine protease]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 14:01:~~04 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:49:11 2008''

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@@ Line 1: / Line 1: @@
-[[Image:2sfa.jpg|left|200px]]<br /><applet load="2sfa" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:2sfa.jpg|left|200px]]<br /><applet load="2sfa" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="2sfa, resolution 1.6&Aring;" />
 '''SERINE PROTEINASE FROM STREPTOMYCES FRADIAE ATCC 14544'''<br />
 ==Overview==
-A proteinase having wide substrate specificity was isolated from, Streptomyces fradiae ATCC 14544. This proteinase, which we propose to call, SFase-2, was purified from the culture filtrate by S-Sepharose, chromatography. The purified enzyme showed an apparent molecular mass of, 19 kDa on SDS/PAGE. When synthetic peptides were used as substrates, SFase-2 showed broad substrate specificity. It also hydrolyzed keratin, elastin and collagen as proteinaceous substrates. It was completely, inhibited by diisopropylfluorophosphate and chymostatin, but not by, tosylphenylalaninechloromethane, tosyllysinechloromethane or EDTA, indicating that it can be classified as a serine proteinase. The matured, protein sequence of SFase-2 was determined by a combination of amino acid, sequencing and the DNA sequencing of the gene. SFase-2, consisting of 191, amino acids, is a novel proteinase. It showed 76% similarity in the amino, acid sequence with Streptomyces griseus proteinase A [Johnson P. and, Smillie L. B. (1974) FEBS Lett. 47, 1-6]. For insight into the, three-dimensional structure of SFase-2, we obtained single crystals by the, vapor diffusion method using sodium phosphate as a precipitant. These, crystals belonged to the orthorhombic, space group P2(1)2(1)2(1) with cell, dimensions a = 6.92 nm, b = 7.28 nm, c = 2.99 nm; one molecule was present, in the asymmetric unit.
+A proteinase having wide substrate specificity was isolated from Streptomyces fradiae ATCC 14544. This proteinase, which we propose to call SFase-2, was purified from the culture filtrate by S-Sepharose chromatography. The purified enzyme showed an apparent molecular mass of 19 kDa on SDS/PAGE. When synthetic peptides were used as substrates, SFase-2 showed broad substrate specificity. It also hydrolyzed keratin, elastin and collagen as proteinaceous substrates. It was completely inhibited by diisopropylfluorophosphate and chymostatin, but not by tosylphenylalaninechloromethane, tosyllysinechloromethane or EDTA, indicating that it can be classified as a serine proteinase. The matured protein sequence of SFase-2 was determined by a combination of amino acid sequencing and the DNA sequencing of the gene. SFase-2, consisting of 191 amino acids, is a novel proteinase. It showed 76% similarity in the amino acid sequence with Streptomyces griseus proteinase A [Johnson P. and Smillie L. B. (1974) FEBS Lett. 47, 1-6]. For insight into the three-dimensional structure of SFase-2, we obtained single crystals by the vapor diffusion method using sodium phosphate as a precipitant. These crystals belonged to the orthorhombic, space group P2(1)2(1)2(1) with cell dimensions a = 6.92 nm, b = 7.28 nm, c = 2.99 nm; one molecule was present in the asymmetric unit.
 ==About this Structure==
-SFA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_fradiae Streptomyces fradiae]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2SFA OCA].
+SFA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_fradiae Streptomyces fradiae]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2SFA OCA].
 ==Reference==
@@ Line 18: / Line 18: @@
 [[Category: serine protease]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 14:01:04 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:49:11 2008''