Identification and Characterization of Cellulose-Binding Domains in Xylanase A of Clostridium stercorarium

ABSTRACT The gene (chi92) encoding the extracellular chitinase of Aeromonas hydrophila JP101 has been cloned and expressed in Escherichia coli. The mature form of Chi92 is an 842-amino-acid (89.830-kDa) modular enzyme comprised of a family 18 catalytic domain, an unknown-function region (the A region), and three chitin-binding domains (ChBDs; Chi92-N, ChBDCI, and ChBDCII). The C-terminally repeated ChBDs, ChBDCI and ChBDCII, were grouped into family V of cellulose-binding domains on the basis of sequence homology. Chitin binding and enzyme activity studies with C-terminally truncated Chi92 derivatives lacking ChBDs demonstrated that the ChBDs are responsible for its adhesion to unprocessed and colloidal chitins. Further adsorption experiments with glutathione S-transferase (GST) fusion proteins (GST-CI and GST-CICII) demonstrated that a single ChBD (ChBDCI) could promote efficient chitin and cellulose binding. In contrast to the two C-terminal ChBDs, the Chi92-N domain is similar to ChiN of Serratia marcescens ChiA, which has been proposed to participate in chitin binding. A truncated derivative of Chi92 that contained only a catalytic domain and Chi92-N still exhibited insoluble-chitin-binding and hydrolytic activities. Thus, it appears that Chi92 contains Chi92-N as the third ChBD in addition to two ChBDs (ChBDCI and ChBDCII).

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Characterization of hybrid proteins consisting of the catalytic domains of Clostridium and Ruminococcus endoglucanases, fused to Pseudomonas non-catalytic cellulose-binding domains

Biochemical Journal ◽

10.1042/bj2790787 ◽

1991 ◽

Vol 279 (3) ◽

pp. 787-792 ◽

Cited By ~ 22

Author(s):

D M Poole ◽

A J Durrant ◽

G P Hazlewood ◽

H J Gilbert

Keyword(s):

Catalytic Domain ◽

Binding Capacity ◽

Binding Domains ◽

Hybrid Proteins ◽

C Terminus ◽

N Terminus ◽

Fusion Enzymes ◽

Cellulose Binding

The N-terminal 160 or 267 residues of xylanase A from Pseudomonas fluorescens subsp. cellulosa, containing a non-catalytic cellulose-binding domain (CBD), were fused to the N-terminus of the catalytic domain of endoglucanase E (EGE') from Clostridium thermocellum. A further hybrid enzyme was constructed consisting of the 347 N-terminal residues of xylanase C (XYLC) from P. fluorescens subsp. cellulosa, which also constitutes a CBD, fused to the N-terminus of endoglucanase A (EGA) from Ruminococcus albus. The three hybrid enzymes bound to insoluble cellulose, and could be eluted such that cellulose-binding capacity and catalytic activity were retained. The catalytic properties of the fusion enzymes were similar to EGE' and EGA respectively. Residues 37-347 and 34-347 of XYLC were fused to the C-terminus of EGE' and the 10 amino acids encoded by the multiple cloning sequence of pMTL22p respectively. The two hybrid proteins did not bind cellulose, although residues 39-139 of XYLC were shown previously to constitute a functional CBD. The putative role of the P. fluorescens subsp. cellulosa CBD in cellulase action is discussed.

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Substrate-binding domains of glycanases from Streptomyces lividans: characterization of a new family of xylan-binding domains

Biochemical Journal ◽

10.1042/bj3300041 ◽

1998 ◽

Vol 330 (1) ◽

pp. 41-45 ◽

Cited By ~ 36

Author(s):

Claude DUPONT ◽

Martin ROBERGE ◽

François SHARECK ◽

Rolf MOROSOLI ◽

Dieter KLUEPFEL

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Substrate Binding ◽

Streptomyces Lividans ◽

Cellulose Binding Domain ◽

Binding Domains ◽

New Family ◽

Cellulose Binding ◽

Sequence Similarities

The substrate-binding domains of six glycanases from Streptomyces lividans were investigated to determine their specificity towards cellulose and xylan. Based upon amino acid sequence similarities, four of the six domains could be assigned to existing cellulose-binding domain families. However, the binding domains of xylanase A and arabinofuranosidase B could not be classified in any of the known families and should therefore be classified as members of a new family. Evidence is also presented that this new family is one of true xylan-binding domains.

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Identification and characterization of single-stranded-DNA-binding proteins from Thermus thermophilus and Thermus aquaticus – new arrangement of binding domains b bThe GenBank accession numbers for the sequences reported in this paper are AF079160 and AF276705.

Microbiology ◽

10.1099/00221287-148-10-3307 ◽

2002 ◽

Vol 148 (10) ◽

pp. 3307-3315 ◽

Cited By ~ 43

Author(s):

Sławomir Dąbrowski ◽

Marcin Olszewski ◽

Rafał Piątek ◽

Anna Brillowska-Dąbrowska ◽

Grażyna Konopa ◽

...

Keyword(s):

Dna Binding ◽

Binding Proteins ◽

Thermus Thermophilus ◽

Dna Binding Proteins ◽

Thermus Aquaticus ◽

Single Stranded Dna ◽

Binding Domains ◽

Identification And Characterization

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Identification and Characterization of a Cellulose Binding Heptapeptide Revealed by Phage Display

Biomacromolecules ◽

10.1021/bm4001876 ◽

2013 ◽

Vol 14 (6) ◽

pp. 1795-1805 ◽

Cited By ~ 21

Author(s):

Jing Guo ◽

Jeffrey M. Catchmark ◽

Mohamed Naseer Ali Mohamed ◽

Alan James Benesi ◽

Ming Tien ◽

...

Keyword(s):

Phage Display ◽

Cellulose Binding ◽

Identification And Characterization

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Identification and characterization of a previously undescribed family of sequence-specific DNA-binding domains

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1221734110 ◽

2013 ◽

Vol 110 (19) ◽

pp. 7660-7665 ◽

Cited By ~ 48

Author(s):

M. B. Lohse ◽

A. D. Hernday ◽

P. M. Fordyce ◽

L. Noiman ◽

T. R. Sorrells ◽

...

Keyword(s):

Dna Binding ◽

Dna Binding Domains ◽

Binding Domains ◽

Identification And Characterization

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Identification and characterization of multiple novel Rab–myosin Va interactions

Molecular Biology of the Cell ◽

10.1091/mbc.e13-05-0236 ◽

2013 ◽

Vol 24 (21) ◽

pp. 3420-3434 ◽

Cited By ~ 62

Author(s):

Andrew J. Lindsay ◽

Florence Jollivet ◽

Conor P. Horgan ◽

Amir R. Khan ◽

Graça Raposo ◽

...

Keyword(s):

Rab Gtpase ◽

Binding Domains ◽

Central Stalk ◽

Myosin Va ◽

Alternatively Spliced ◽

Total Pool ◽

Novel Interactions ◽

Identification And Characterization ◽

Two Hybrid

Myosin Va is a widely expressed actin-based motor protein that binds members of the Rab GTPase family (3A, 8A, 10, 11A, 27A) and is implicated in many intracellular trafficking processes. To our knowledge, myosin Va has not been tested in a systematic screen for interactions with the entire Rab GTPase family. To that end, we report a yeast two-hybrid screen of all human Rabs for myosin Va-binding ability and reveal 10 novel interactions (3B, 3C, 3D, 6A, 6A′, 6B, 11B, 14, 25, 39B), which include interactions with three new Rab subfamilies (Rab6, Rab14, Rab39B). Of interest, myosin Va interacts with only a subset of the Rabs associated with the endocytic recycling and post-Golgi secretory systems. We demonstrate that myosin Va has three distinct Rab-binding domains on disparate regions of the motor (central stalk, an alternatively spliced exon, and the globular tail). Although the total pool of myosin Va is shared by several Rabs, Rab10 and Rab11 appear to be the major determinants of its recruitment to intracellular membranes. We also present evidence that myosin Va is necessary for maintaining a peripheral distribution of Rab11- and Rab14-positive endosomes.

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