#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

The HMW1C Protein Is a Glycosyltransferase That Transfers Hexose Residues to Asparagine Sites in the HMW1 Adhesin


The Haemophilus influenzae HMW1 adhesin is a high-molecular weight protein that is secreted by the bacterial two-partner secretion pathway and mediates adherence to respiratory epithelium, an essential early step in the pathogenesis of H. influenzae disease. In recent work, we discovered that HMW1 is a glycoprotein and undergoes N-linked glycosylation at multiple asparagine residues with simple hexose units rather than N-acetylated hexose units, revealing an unusual N-glycosidic linkage and suggesting a new glycosyltransferase activity. Glycosylation protects HMW1 against premature degradation during the process of secretion and facilitates HMW1 tethering to the bacterial surface, a prerequisite for HMW1-mediated adherence. In the current study, we establish that the enzyme responsible for glycosylation of HMW1 is a protein called HMW1C, which is encoded by the hmw1 gene cluster and shares homology with a group of bacterial proteins that are generally associated with two-partner secretion systems. In addition, we demonstrate that HMW1C is capable of transferring glucose and galactose to HMW1 and is also able to generate hexose-hexose bonds. Our results define a new family of bacterial glycosyltransferases.


Vyšlo v časopise: The HMW1C Protein Is a Glycosyltransferase That Transfers Hexose Residues to Asparagine Sites in the HMW1 Adhesin. PLoS Pathog 6(5): e32767. doi:10.1371/journal.ppat.1000919
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000919

Souhrn

The Haemophilus influenzae HMW1 adhesin is a high-molecular weight protein that is secreted by the bacterial two-partner secretion pathway and mediates adherence to respiratory epithelium, an essential early step in the pathogenesis of H. influenzae disease. In recent work, we discovered that HMW1 is a glycoprotein and undergoes N-linked glycosylation at multiple asparagine residues with simple hexose units rather than N-acetylated hexose units, revealing an unusual N-glycosidic linkage and suggesting a new glycosyltransferase activity. Glycosylation protects HMW1 against premature degradation during the process of secretion and facilitates HMW1 tethering to the bacterial surface, a prerequisite for HMW1-mediated adherence. In the current study, we establish that the enzyme responsible for glycosylation of HMW1 is a protein called HMW1C, which is encoded by the hmw1 gene cluster and shares homology with a group of bacterial proteins that are generally associated with two-partner secretion systems. In addition, we demonstrate that HMW1C is capable of transferring glucose and galactose to HMW1 and is also able to generate hexose-hexose bonds. Our results define a new family of bacterial glycosyltransferases.


Zdroje

1. BenzI

SchmidtMA

2001 Glycosylation with heptose residues mediated by the aah gene product is essential for adherence of the AIDA-1 adhesin. Mol Microbiol 40 1403 1413

2. BrimerC

MontieT

1998 Cloning and comparison of fliC genes and identification of glycosylation in the flagellin of Pseudomonas aeruginosa a-type strains. J Bacteriol 180 3209 3217

3. CastricP

1995 pilO, a gene required for glycosylation of Pseudomonas aeruginosa 1244 pilin. Microbiology 141 1247 1254

4. DoigP

KinsellaN

GuerryP

TrustTJ

1996 Characterization of a post-translational modification of Campylobacter flagellin: identification of a sero-specific glycosyl moiety. Mol Microbiol 19 379 387

5. LindenthalC

ElsinghorstEA

1999 Identification of a glycoprotein produced by enterotoxigenic Escherichia coli. Infect Immun 67 4084 4091

6. SchmidtMA

RileyLW

BenzI

2003 Sweet new world: glycoproteins in bacterial pathogens. Trends Microbiol 11 554 561

7. SzymanskiCM

YaoR

EwingCP

TrustTJ

GuerryP

1999 Evidence for a system of general protein glycosylation in Campylobacter jejuni. Mol Microbiol 32 1022 1030

8. SzymanskiCM

WrenBW

2005 Protein glycosylation in bacterial mucosal pathogens. Nature Rev Microbiol 3 225 237

9. VirjiM

1997 Post-translational modifications of meningococcal pili. Identification of common substituents: glycans and α-glycerophosphate – a review. Gene 192 141 147

10. WackerM

LintonD

HitchenPG

Nita-LazarM

HaslamSM

2002 N-linked glycosylation in Campylobacter jejuni and its functional transfer into E. coli. Science 298 1790 1793

11. TurkDC

1984 The pathogenicity of Haemophilus influenzae. J Med Microbiol 18 1 16

12. St GemeJWIII

1993 Nontypeable Haemophilus influenzae disease: epidemiology, pathogenesis, and prospects for prevention. Infect Agents Dis 2 1 16

13. St GemeJWIII

FalkowS

BarenkampSJ

1993 High-molecular-weight proteins of nontypeable Haemophilus influenzae mediate attachment to human epithelial cells. Proc Natl Acad Sci, USA 90 2875 2879

14. St GemeJWIII

GrassS

1998 Secretion of the Haemophilus influenzae HMW1 and HMW2 adhesins involves a periplasmic intermediate and requires the HMWB and HMWC proteins. Mol Microbiol 27 617 630

15. BarenkampSJ

St GemeJWIII

1994 Genes encoding high molecular weight adhesion proteins of nontypeable Haemophilus influenzae are part of gene clusters. Infect Immun 62 3320 3328

16. BuscherAZ

BurmeisterK

BarenkampSJ

St GemeJWIII

2004 Evolutionary and functional relationships among the nontypeable Haemophilus influenzae HMW family of adhesins. J Bacteriol 186 4209 4217

17. Jacob-DubuissonF

LochtC

AntoineR

2001 Two-partner secretion in Gram-negative bacteria: a thrifty, specific pathway for large virulence proteins. Mol Microbiol 40 306 313

18. GrassS

St GemeJWIII

2000 Maturation and secretion of the non-typable Haemophilus influenzae HMW1 adhesin: roles of the N-terminal and C-terminal domains. Mol Microbiol 36 55 67

19. St GemeJWIII

KumarVV

CutterD

BarenkampSJ

1998 Prevalence and distribution of the hmw and hia genes and the HMW and Hia adhesins among genetically diverse strains of nontypeable Haemophilus influenzae. Infect Immun 66 364 368

20. BarenkampSJ

LeiningerE

1992 Cloning, expression, and DNA sequence analysis of genes encoding nontypeable Haemophilus influenzae high-molecular-weight proteins related to filamentous hemagglutinin of Bordetella pertussis. Infect Immun 60 1302 1313

21. SuranaNK

GrassS

HardyGG

HuilinL

ThanassiDG

St. GemeJWIII

2004 Evidence for conservation of architecture and physical properties of Omp85-like proteins throughout evolution. Proc Natl Acad Sci USA 101 14497 14502

22. GrassS

2003 The Haemophilus influenzae HMW1 adhesin is glycosylated in a process that requires HMW1C and phosphoglucomutase, an enzyme involved in lipooligosaccharide biosynthesis. Mol Microbiol 48 737 751

23. GrossJ

BuscherAZ

SwordsWE

ApicellaMA

BarenkampSJ

2008 The Haemophilus influenzae HMW1 adhesin is glycosylated with an unusual N-linked carbohydrate modification. J Biol Chem 283 26010 26015

24. AndersonP

JohnstonRBJr

SmithDH

1972 Human serum activity against Haemophilus influenzae type b. J Clin Invest 51 31 38

25. KelleherDJ

GilmoreR

2006 An evolving view of the eukaryotic oligosaccharyltransferase. Glycobiology 16 47R 62R

26. ChabanB

VoisinS

KellyJ

LoganSM

JarrellKF

2006 Identification of genes involved in the biosynthesis and attachment of Methanococcus voltae N-linked glycans: insight into N-linked glycosylation pathways in Archaea. Mol Microbiol 61 259 268

27. Abu-QarnM

Yurist-DoutschS

GiordanoA

TraunerA

MorrisHR

2007 Haloferax volcanii AglB and AglD are involved in N-glycosylation of the S-layer glycoprotein and proper assembly of the surface layer. J Mol Biol 374 1224 1236

28. FleckensteinJM

RoyK

FischerJF

BurkittM

2006 Identification of a two-partner secretion locus. Infect Immun 74 2245 2258

29. RoyK

HamiltonD

AllenKP

RandolphMP

FleckensteinJM

2008 The EtpA exoprotein of enterotoxigenic Escherichia coli promotes intestinal colonization and is a protective antigen in an experimental model of murine infection. Infect Immun 76 2106 2112

30. HoodDW

DeadmanME

AllenT

MasoudH

MartinA

2006 Use of the complete genome sequence information of Haemophilus influenzae strain Rd to investigate lipopolysaccharide biosynthesis. Mol Microbiol 22 951 965

31. SambrookJ

RusselDW

2001 Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, New York

32. HerriottRM

MeyerEM

VogtM

1970 Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol 101 517 524

33. BuscherAZ

GrassS

HeuserJ

RothR

St GemeJWIII

2006 Surface anchoring of a bacterial adhesin secreted by the two-partner secretion pathway. Mol Microbiol 61 470 83

34. MedzihradszkyKF

CampbellJM

BaldwinMA

FalickAM

JuhaszP

2000 The characteristics of peptide collision-induced dissociation using a high-performance MALDI-TOF/TOF tandem mass spectrometer. Anal Chem 72 552 558

35. NittisT

GuittatL

LeDucRD

DaoB

DuxinJB

2010 Revealing novel telomere proteins using in vivo crosslinking, tandem affinity purification and label-free quantitative LC-FTICR-MS. Mol Cell Proteomics Jan 22, 2010 (Epub ahead of print)

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2010 Číslo 5
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#