ProGP101

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ProGP ID ProGP101
Validation Status Characterized
Organism Information
Organism NamePaenibacillus (Bacillus) macerans and Bacillus amyloliquefaciens (velezensis)
Domain Bacteria
Classification Family: Bacillaceae
Order: Bacillales
Class: Bacilli (or Firmibacteria)
Division or phylum: "Firmicutes"
Taxonomic ID (NCBI) 44252
Genome Sequence(s)
EMBL X55959
Protein Information
Protein NameH(A16-M) (1,3-1,4)-beta-glucanase
UniProtKB/SwissProt ID P23904
EMBL-CDSCAA39426.1
UniProtKB Sequence >sp|P23904|GUB_PAEMA Beta-glucanase OS=Paenibacillus macerans PE=1 SV=2 MKKKSCFTLVTTFAFSLIFSVSALAGSVFWEPLSYFNRSTWEKADGYSNGGVFNCTWRAN NVNFTNDGKLKLGLTSSAYNKFDCAEYRSTNIYGYGLYEVSMKPAKNTGIVSSFFTYTGP AHGTQWDEIDIEFLGKDTTKVQFNYYTNGVGGHEKVISLGFDASKGFHTYAFDWQPGYIK WYVDGVLKHTATANIPSTPGKIMMNLWNGTGVDDWLGSYNGANPLYAEYDWVKYTSN
Sequence length 237 AA
Subcellular LocationSecreted
Function It is the hybrid enzyme that catalyses cleavage of (1,4)-beta-linkages of O-substituted beta-D-glucanopyranosyl residues.
Protein Structure
PDB ID 1CPM, 1GLH, 2AYH
Glycosylation Status
Glycosylation Type N (Asn) linked
Experimentally Validated Glycosite(s) in Full Length Protein(Signal peptide: 1-25) N56, N210
Experimentally Validated Glycosite(s ) in Mature ProteinN31, N185
Glycosite(s) Annotated Protein Sequence MKKKSCFTLVTTFAFSLIFSVSALAQTGGSFFEPFNSYNSGTWEKADGYSNGGVFN*(56)CTWR ANNVNFTNDGKLKLGLTSSAYNKFDCAEYRSTNIYGYGLYEVSMKPAKNTGIVSSFFTYT GPAHGTQWDEIDIEFLGKDTTKVQFNYYTNGVGGHEKVISLGFDASKGFHTYAFDWQPGY IKWYVDGVLKHTATANIPSTPGKIMMNLWN*(210)GTGVDDWLGSYNGANPLYAEYDWVKYTSN
This sequence has 16 N-terminal amino acids (colored red) derived from AMY. The remaining residues are from MAC.
Sequence Around Glycosites (21 AA) ADGYSNGGVFNCTWRANNVNF
TPGKIMMNLWNGTGVDDWLGS
Glycosite Sequence Logo
Technique(s) used for Glycosylation DetectionMass shift detected after deglycosylation with Endo Hf and PNGase F
Technique(s) used for Glycosylated Residue(s) Detection N-terminal sequencing
Protein Glycosylation- Implication There is a large influence of N glycosylation on the thermostability of the hybrid enzyme. 16- and 133-fold decrease of thermostability has been observed after treatment with endoglycosidase H and peptide:N-glycosidase F (that remove glycans). This indicates that N-glycans are a major determinant for the resistance of this hybrid glucanase to thermal inactivation.
Glycan Information
Glycan Annotation Linkage: GlcNAc-N-Asn
Literature
Additional CommentEngineered glycoprotein
The glucanases from which the hybrid enzyme is made, are not glycosylated in their native hosts. Hybrid enzyme containing 16 N-terminal amino acids from Bacillus amyloliquefaciens (1,3-1,4)-beta-glucanase (AMY), and remaining C-terminal part and signal peptide from B. macerans (1,3-1,4)-beta-glucanase (MAC), was expressed in Saccharomyces cerevisiae to see the effect of N-glycosylation on the thermostability of the hybrid enzyme.
Information from UniProtKB and EMBL has been provided for MAC enzyme as it constitutes the major part of the hybrid enzyme (239 AA). UniProtKB ID for AMY is P07980.
Year of Identification1994
Year of Identification Month Wise1994.01.01
Year of Validation 1994
Reference Olsen, O., Thomesen, K. K. (1991) Improvement of bacterial P-glucanase thermostability by glycosylation. J Gen Microbiol, 137, 579–585.
AuthorOlsen, O., Thomesen, K. K.
Research GroupCarlsberg Laboratory, Department of Physiology, Gamle Carlsbergvej 10, DK-2500 Copenhagen Valby, Denmark
Corresponding Author Thomesen, K. K.
ContactCarlsberg Laboratory, Department of Physiology, Gamle Carlsbergvej 10, DK-2500 Copenhagen Valby, Denmark
Reference1) Hahn, M., Keitel, T. and Heinemann, U. (1995) Crystal and molecular structure at 0.16-nm resolution of the hybrid Bacillus endo-1,3-1,4-beta-D-glucan 4-glucanohydrolase H(A16-M). Eur J Biochem, 232, 849-858. [PubMed: 7588726]
AuthorHahn, M., Keitel, T. and Heinemann, U.
Research GroupForschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Corresponding Author Heinemann, U
ContactForschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Reference Hahn, M., Piotukh, K., Borriss, R. and Heinemann, U. (1994) Native-like in vivo folding of a circularly permuted jellyroll protein shown by crystal structure analysis. Proc Natl Acad Sci U S A, 91, 10417-10421. [PubMed: 7937966]
Author Hahn, M., Piotukh, K., Borriss, R. Heinemann, U.
Research GroupMax-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Corresponding Author Heinemann, U.
ContactMax-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Reference Meldgaard, M. and Svendsen, I. (1994) Different effects of N-glycosylation on the thermostability of highly homologous bacterial (1,3-1,4)-beta-glucanases secreted from yeast. Microbiology, 140 ( Pt 1), 159-166. [PubMed: 8162185]
Author Meldgaard, M. Svendsen, I.
Research GroupDepartment of Physiology, Carlsberg Laboratory, Copenhagen, Denmark.
Corresponding Author Svendsen, I.
ContactDepartment of Physiology, Carlsberg Laboratory, Copenhagen, Denmark.
ReferenceMeldgaard, M. and Svendsen, I. (1994) Different effects of N-glycosylation on the thermostability of highly homologous bacterial (1,3-1,4)-beta-glucanases secreted from yeast. Microbiology, 140 ( Pt 1), 159-166. [PubMed: 8162185]
Author Svendsen, I.
Research GroupDepartment of Physiology, Carlsberg Laboratory, Copenhagen, Denmark.
Corresponding Author Meldgaard, M. Svendsen, I.
ContactDepartment of Physiology, Carlsberg Laboratory, Copenhagen, Denmark.
Reference Keitel, T., Meldgaard, M. and Heinemann, U. (1994) Cation binding to a Bacillus (1,3-1,4)-beta-glucanase. Geometry, affinity and effect on protein stability. Eur J Biochem, 222, 203-214. [PubMed: 8200344]
Author Heinemann, U.
Research GroupInstitut für Kristallographie, Freie Universität Berlin, Germany
Corresponding Author Keitel, T., Meldgaard, M. Heinemann, U.
ContactInstitut für Kristallographie, Freie Universität Berlin, Germany