Glycan Array Experiment: primscreen_6593
Glycan Array ID
:
primscreen_6593
Sample Name
:
Streptosporangium roseum F-type lectin a-L-fucosidase
Sample Category
:
Plant Lectins
Investigator:
:
Ramya TNC
Download Data:
Sample Information:
Name
:
Streptosporangium roseum F-type lectin a-L-fucosidase
Complete Name
:
S. roseum fucosidase, S. roseum FLD
Family
:
Viral / Bacterial Lectin
Sub Family
:
F-type lectin/ CBM47
Gene Symbol
:
Information not entered/not applicable.
Synonyms
:
Information not entered/not applicable.
Species Common Name:
:
Information not entered/not applicable.
Species Scientific Name
:
Streptosporangium roseum
Estimated Purity
:
>85% purity, contaminants are expected to be E. coli BL21-DE3 proteins
Mutation/Chimera
:
C-ter hexahistidine tagged wildtype full length or separate domains, mutations in FLD and/or fucosidase domain
Entry Date [yyyy-mm-dd]
:
2014-04-11
Status
:
Private
Primary Sequence
:
MRRKLLLVPAVGLLVIGSIFATQPVASADTPPVYEPTVESLNSHPVPQWFNDDKFGIFIHWGAYSVPAWGPRGSYAEWYGNYMNAGGSATNAHHKATYGQDFNYDAFLQQWKAEKFDPADWVKLFKDAGAKYFVLTSKHHEGMALWDSKSSGRDSVDLGPGRDLAKELFDAARKDEAKLKAGFYYSLYEWYNPAYTGRPATNPYTGAEIPYTGAPGGGDYVKDYMLPQMRELIDGYDPDIIWCDGQWEKPASYWNTAGVIADYYNKALNSGKEVAVANRCKIQSGNLDSPELDFQTPEYTVKPDIDPVKWESSRGIAHSYGYNQNEPEEDHLTSDQLVDSLVDIVSKNGNLLLDIGPKADGTIPEIQRQRLLDIGAWLKANGEAIYGTTYWNRAEEKGGPDDVRYTAKDATLYATALKWPGEQLTLGADLPVDHSTRITMLGSGARLSWSRDEQGRVVVNTPQEAGKHAYVFKIETPGVRSLLRTSSSLAKEIAPGRTISGELTVTNPGKRHTPATKLSLTVPQGWTATLGAPRVRPLGPGESVKVPISVSAPEGVAPAPYTLGLYQRTGRMGTTTALPLTVNRPNLSLGKPATQKSTGYDAPASRAVDGNTGGDWSAGSTTHTAEPEKQAWWQVDLGASARLDSVDVWNRLDCCADRLKDFWVMASDQPFTTDDLDQARTAPGVTAVHVGEQAGSPSKVKLPEGTRGRYVRIQLASPSNPLSLAEVQVRGSrFLD(proteinwithjustF-typelectindomain)MARPNLSLGKPATQKSTGYDAPASRAVDGNTGGDWSAGSTTHTAEPEKQAWWQVDLGASARLDSVDVWNRLDCCADRLKDFWVMASDQPFTTDDLDQARTAPGVTAVHVGEQAGSPSKVKLPEGTRGRYVRIQLASPSNPLSLAEVQVRLEHHHHHHSrNPCBM(proteinwithNPCBM-associateddomain)MAVRSLLRTSSSLAKEIAPGRTISGELTVTNPGKRHTPATKLSLTVPQGWTATLGAPRVRPLGPGESVKVPISVSAPEGVAPAPYTLGLYQRTGRMGTTTALPLTVNLEHHHHHH
Comments
:
Hemagglutination for confirming lectin activity
A 2% suspension of human type-O erythrocytes is made in 20 mM Tris HCl, 10 mM CaCl2 0.15 M NaCl, pH 7.5
25 microlitres of the erythrocyte suspension is mixed with 25 microlitres of the recombinant protein solution in a U-bottomed microtitre plate and incubated at 37 C
Hemagglutination is observed after 1 hour
Known Sites of Expression
:
The gene encoding full length or separate F-type lectin domain or NPCBM associated domain of Streptosporangium roseum fucosidase without signal peptide and stop codon was cloned within NcoI and XhoI sites in pET28a(+) to enable expression of C-terminal His tagged protein. Cultures were grown till OD of 0.8, induced with 0.1 mM IPTG at 20 C for 10 hours, and then protein was purified from lysate by metal ion affinity chromatography. Lectin activity was confirmed by hemagglutination assay using type O erythrocytes.
External References :
Swissprot
:
D2B1P1_STRRD
Genbank
:
ACZ87343.1
Experiment Information:
Status:
:
Public
Title/Project Description:
:
The purpose of this request is to perform the analysis of Streptosporangium roseum fucosidase protein containing a fucosidase domain, an NPCBM domain and an F-type lectin domain, on the glycan array.
Associated Resource Requests:
:
cfg_rRequest_3001
Glycan Array Version:
:
PA_v52
Protocol Used:
:
Protocol Tertiary step
Replicate Number:
:
1
Result Nature:
:
Data
Protein/Sample Concentration:
:
100 μg/ml
Experiment Date [yyyy-mm-dd]:
:
2014-08-08
Supplying Investigator
:
Ramya TNC
Specific Assay Information
:
Sample was diluted in binding buffer containing 10 mM CaCl2 and was detected with anti-His (Sigma, sent by PI, 1:1000) followed by detected with anti-rabbit IgG-488 (Invitrogen, 5 ug/ml).
Annotation
:
WT FTP
Protein Sample Analyzed
:
Streptosporangium roseum F-type lectin a-L-fucosidase