scholarly journals Cholesterol 3-Sulfate Interferes with Cornified Envelope Assembly by Diverting Transglutaminase 1 Activity from the Formation of Cross-links and Esters to the Hydrolysis of Glutamine

2000 ◽  
Vol 275 (4) ◽  
pp. 2636-2646 ◽  
Author(s):  
Zoltán Nemes ◽  
Máté Demény ◽  
Lyuben N. Marekov ◽  
László Fésüs ◽  
Peter M. Steinert
Keyword(s):  
Cornified Envelope ◽  
Transglutaminase 1 ◽  
Cross Links ◽  
Hydrolysis Of

scholarly journals Lipid Metabolic Events Underlying the Formation of the Corneocyte Lipid Envelope

2021 ◽  
pp. 1-13
Author(s):  
Philip W. Wertz
Keyword(s):  
Stratum Corneum ◽  
Outer Surface ◽  
Barrier Function ◽  
Skin Surface ◽  
Schiff’S Base ◽  
Cornified Envelope ◽  
Epoxy Alcohol ◽  
Viable Epidermis ◽  
Transglutaminase 1 ◽  
Base Formation

Cornified cells of the stratum corneum have a monolayer of an unusual lipid covalently attached to the outer surface. This is referred to as the corneocyte lipid envelope (CLE). It consists of a monolayer of ω-hydroxyceramides covalently attached to the outer surface of the cornified envelope. The CLE is essential for proper barrier function of the skin and is derived from linoleate-rich acylglucosylceramides synthesized in the viable epidermis. Biosynthesis of acylglucosylceramide and its conversion to the cornified envelope is complex. Acylglucosylceramide in the bounding membrane of the lamellar granule is the precursor of the CLE. The acylglucosylceramide in the limiting membrane of the lamellar granule may be oriented with the glucosyl moiety on the inside. Conversion of the acylglucosylceramide to the CLE requires removal of the glucose by action of a glucocerebrosidase. The ester-linked fatty acid may be removed by an as yet unidentified esterase, and the resulting ω-hydroxyceramide may become ester linked to the outer surface of the cornified envelope through action of transglutaminase 1. Prior to removal of ester-linked fatty acids, linoleate is oxidized to an epoxy alcohol through action of 2 lipoxygenases. This can be further oxidized to an epoxy-enone, which can spontaneously attach to the cornified envelope through Schiff’s base formation. Mutations of genes coding for enzymes involved in biosynthesis of the CLE result in ichthyosis, often accompanied by neurologic dysfunction. The CLE is recognized as essential for barrier function of skin, but many questions about details of this essentiality remain. What are the relative roles of the 2 mechanisms of lipid attachment? What is the orientation of acylglucosylceramide in the bounding membrane of lamellar granules? Some evidence supports a role for CLE as a scaffold upon which intercellular lamellae unfold, but other evidence does not support this role. There is also controversial evidence for a role in stratum corneum cohesion. Evidence is presented to suggest that covalently bound ω-hydroxyceramides serve as a reservoir for free sphingosine that can serve in communicating with the viable epidermis and act as a potent broad-acting antimicrobial at the skin surface. Many questions remain.

scholarly journals Localization of the Transglutaminase Cross-Linking Sites in the Bacillus subtilis Spore Coat Protein GerQ

2006 ◽  
Vol 188 (21) ◽  
pp. 7609-7616 ◽  
Author(s):  
Alicia Monroe ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Coat Protein ◽  
Cross Linking ◽  
Spore Coat ◽  
Bacillus Subtilis Spore ◽  
Binding Partners ◽  
Lysine Residues ◽  
Cross Links ◽  
Hydrolysis Of ◽  
Spore Coat Protein

ABSTRACT The Bacillus subtilis spore coat protein GerQ is necessary for the proper localization of CwlJ, an enzyme important in the hydrolysis of the peptidoglycan cortex during spore germination. GerQ is cross-linked into high-molecular-mass complexes in the spore coat late in sporulation, and this cross-linking is largely due to a transglutaminase. This enzyme forms an ε-(γ-glutamyl) lysine isopeptide bond between a lysine donor from one protein and a glutamine acceptor from another protein. In the current work, we have identified the residues in GerQ that are essential for transglutaminase-mediated cross-linking. We show that GerQ is a lysine donor and that any one of three lysine residues near the amino terminus of the protein (K2, K4, or K5) is necessary to form cross-links with binding partners in the spore coat. This leads to the conclusion that all Tgl-dependent GerQ cross-linking takes place via these three lysine residues. However, while the presence of any of these three lysine residues is essential for GerQ cross-linking, they are not essential for the function of GerQ in CwlJ localization.

scholarly journals Peptidoglycan Cross-Linking in Glycopeptide-Resistant Actinomycetales

2014 ◽  
Vol 58 (3) ◽  
pp. 1749-1756 ◽  
Author(s):  
Jean-Emmanuel Hugonnet ◽  
Nabila Haddache ◽  
Carole Veckerlé ◽  
Lionel Dubost ◽  
Arul Marie ◽  
...  
Keyword(s):  
Streptomyces Coelicolor ◽  
Residual Activity ◽  
Resistance Mechanisms ◽  
Strain Atcc ◽  
Glycopeptide Resistance ◽  
Content Type ◽  
Peptidoglycan Structure ◽  
Cross Links ◽  
Acyl Donors ◽  
Hydrolysis Of

ABSTRACTSynthesis of peptidoglycan precursors ending ind-lactate (d-Lac) is thought to be responsible for glycopeptide resistance in members of the orderActinomycetalesthat produce these drugs and in related soil bacteria. More recently, the peptidoglycan of several members of the orderActinomycetaleswas shown to be cross-linked byl,d-transpeptidases that use tetrapeptide acyl donors devoid of the target of glycopeptides. To evaluate the contribution of these resistance mechanisms, we have determined the peptidoglycan structure ofStreptomyces coelicolorA(3)2, which harbors avanHAXgene cluster for the production of precursors ending ind-Lac, andNonomuraeasp. strain ATCC 39727, which is devoid ofvanHAXand produces the glycopeptide A40296. Vancomycin retained residual activity againstS. coelicolorA(3)2 despite efficient incorporation ofd-Lac into cytoplasmic precursors. This was due to ad,d-transpeptidase-catalyzed reaction that generated a stem pentapeptide recognized by glycopeptides by the exchange ofd-Lac ford-Ala and Gly. The contribution ofl,d-transpeptidases to resistance was limited by the supply of tetrapeptide acyl donors, which are essential for the formation of peptidoglycan cross-links by these enzymes. In the absence of a cytoplasmic metallo-d,d-carboxypeptidase, the tetrapeptide substrate was generated by hydrolysis of the C-terminald-Lac residue of the stem pentadepsipeptide in the periplasm in competition with the exchange reaction catalyzed byd,d-transpeptidases. InNonomuraeasp. strain ATCC 39727, the contribution ofl,d-transpeptidases to glycopeptide resistance was limited by the incomplete conversion of pentapeptides into tetrapeptides despite the production of a cytoplasmic metallo-d,d-carboxypeptidase. Since the level of drug production exceeds the level of resistance, we propose thatl,d-transpeptidases merely act as a tolerance mechanism in this bacterium.

scholarly journals p-NN′-phenylenebismaleimide, a specific cross-linking agent for F-actin

1978 ◽  
Vol 175 (3) ◽  
pp. 1023-1032 ◽  
Author(s):  
P Knight ◽  
G Offer
Keyword(s):  
Quantitative Analysis ◽  
Sodium Dodecyl Sulphate ◽  
Actin Filaments ◽  
Sodium Dodecyl ◽  
Cysteine Residue ◽  
Cross Linking ◽  
Maximum Value ◽  
Cross Links ◽  
The Cross ◽  
Hydrolysis Of

Covalent cross-links can be inserted between the subunits of F-actin by using p-NN′-phenylenebismaleimide. Cross-linking reaches its maximum value when one molecule of reagent has reacted with each actin subunit. p-NN′-Phenylenebismaleimide reacts initially with a cysteine residue on one subunit, the slower cross-linking reaction involving a lysine residue on a neighbouring subunit. Hydrolysis of the actin-bound reagent limits the extent of cross-linking. Quantitative analysis of the amounts of cross-linked oligomers seen on polyacrylamide gels containing sodium dodecyl sulphate suggests that neither the binding of the reagent to actin nor the formation of cross-links introduces strain into the structure. The cross-links do not join together different F-actin filaments, and evidence is presented that suggests that the cross-links join subunits of the same long-pitched helix.

scholarly journals P4-160: Transglutaminase 1, Transglutaminase 2 and Transglutaminase-catalyzed cross-links colocalize with the pathological lesions in Alzheimer's disease brain

2008 ◽  
Vol 4 ◽  
pp. T717-T717
Author(s):  
Micha M. Wilhelmus ◽  
Sentini C.S. Grunberg ◽  
John G.J.M. Bol ◽  
Jeroen J.M. Hoozemans ◽  
Annemieke J.M. Rozemuller ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Transglutaminase 2 ◽  
Pathological Lesions ◽  
Transglutaminase 1 ◽  
Cross Links

scholarly journals Peptidoglycan Cross-Linking Activity of L,D-Transpeptidases from Clostridium difficile and Inactivation of These Enzymes by β-Lactams

2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Laetitia Sütterlin ◽  
Zainab Edoo ◽  
Jean-Emmanuel Hugonnet ◽  
Jean-Luc Mainardi ◽  
Michel Arthur
Keyword(s):  
Mass Spectrometry ◽  
Clostridium Difficile ◽  
Cross Linking ◽  
Content Type ◽  
Catalytic Activities ◽  
Cross Links ◽  
Covalent Adducts ◽  
The Stability ◽  
Hydrolysis Of ◽  
Insight Into

ABSTRACT In most bacteria, the essential targets of β-lactam antibiotics are the d , d -transpeptidases that catalyze the last step of peptidoglycan polymerization by forming 4→3 cross-links. The peptidoglycan of Clostridium difficile is unusual since it mainly contains 3→3 cross-links generated by l , d -transpeptidases. To gain insight into the characteristics of C. difficile peptidoglycan cross-linking enzymes, we purified the three putative C. difficile l , d -transpeptidase paralogues Ldt Cd1 , Ldt Cd2 , and Ldt Cd3 , which were previously identified by sequence analysis. The catalytic activities of the three proteins were assayed with a disaccharide-tetrapeptide purified from the C. difficile cell wall. Ldt Cd2 and Ldt Cd3 catalyzed the formation of 3→3 cross-links ( l , d -transpeptidase activity), the hydrolysis of the C-terminal d -Ala residue of the disaccharide-tetrapeptide substrate ( l , d -carboxypeptidase activity), and the exchange of the C-terminal d -Ala for d -Met. Ldt Cd1 displayed only l , d -carboxypeptidase activity. Mass spectrometry analyses indicated that Ldt Cd1 and Ldt Cd2 were acylated by β-lactams belonging to the carbapenem (imipenem, meropenem, and ertapenem), cephalosporin (ceftriaxone), and penicillin (ampicillin) classes. Acylation of Ldt Cd3 by these β-lactams was not detected. The acylation efficacy of Ldt Cd1 and Ldt Cd2 was higher for the carbapenems (480 to 6,600 M −1 s −1 ) than for ampicillin and ceftriaxone (3.9 to 82 M −1 s −1 ). In contrast, the efficacy of the hydrolysis of β-lactams by Ldt Cd1 and Ldt Cd2 was higher for ampicillin and ceftriaxone than for imipenem. These observations indicate that Ldt Cd1 and Ldt Cd2 are inactivated only by β-lactams of the carbapenem class due to a combination of rapid acylation and the stability of the resulting covalent adducts.

scholarly journals Role of endopeptidases in peptidoglycan synthesis mediated by alternative cross-linking enzymes in Escherichia coli

2021 ◽  
Author(s):  
Henri Voedts ◽  
Delphine Dorchêne ◽  
Adam Lodge ◽  
Waldemar Vollmer ◽  
Michel Arthur ◽  
...  
Keyword(s):  
Turgor Pressure ◽  
Cross Linking ◽  
Penicillin Binding Proteins ◽  
Peptidoglycan Synthesis ◽  
Dual Specificity ◽  
Common Motif ◽  
Cross Links ◽  
Hydrolysis Of ◽  
Controlled Hydrolysis

ABSTRACTBacteria resist to the turgor pressure of the cytoplasm through a net-like macromolecule, the peptidoglycan, made of glycan strands connected via peptides cross-linked by penicillin-binding proteins (PBPs). We recently reported the emergence of β-lactam resistance resulting from a bypass of PBPs by the YcbB L,D-transpeptidase (LdtD), which form chemically distinct 3→3 cross-links compared to 4→3 formed by PBPs. Here we show that peptidoglycan expansion requires controlled hydrolysis of cross-links and identify amongst eight endopeptidase paralogues the minimum enzyme complements essential for bacterial growth with 4→3 (MepM) and 3→3 (MepM and MepK) cross-links. Purified Mep endopeptidases unexpectedly displayed a 4→3 and 3→3 dual specificity implying recognition of a common motif in the two cross-link types. Uncoupling of the polymerization of glycan chains from the 4→3 cross-linking reaction was found to facilitate the bypass of PBPs by YcbB. These results illustrate the plasticity of the peptidoglycan polymerization machinery in response to the selective pressure of β-lactams.

Use of a Diimidoester Cross-Linking Reagent to Examine the Submit Structure of Rabbit Muscle Pyruvate Kinase

1972 ◽  
Vol 50 (4) ◽  
pp. 416-422 ◽  
Author(s):  
Gregg E. DaVies ◽  
J. Gordin Kaplan
Keyword(s):  
Pyruvate Kinase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cross Linking ◽  
Rabbit Muscle ◽  
Cross Links ◽  
Primary Amino ◽  
Muscle Pyruvate Kinase ◽  
Dimeric Intermediate ◽  
Hydrolysis Of

Rabbit muscle pyruvate kinase, a tetramer of highly similar or identical subunits, is known to dissociate in urea via a dimeric intermediate. To provide additional evidence regarding the structure of the native oligomer, we have treated pyruvate kinase with dimethyl pimelimidate and examined the yields of cross-linked species resolved by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Of four products resolved, predominating species are dimers and tetramers of the pyruvate kinase subunit, while monomers and trimers are present in lesser amounts. These relative yields are consistent with a dimeric structure of the tetramer.In a typical cross-linking reaction, about 86 of the 148 primary amino groups in the pyruvate kinase tetramer are amidinated. Of 53 moles of cross-linking reagent incorporated per mole of tetramer, 38% have reacted monofunctionally, with hydrolysis of the second imidoester group, and 62% have reacted bifunctionally to form intra- and intersubunit cross-links.

Aging Stability of Neoprene Latex. Relation between Cross-Linking and Hydrolysis of Allylic Chlorine

1955 ◽  
Vol 28 (2) ◽  
pp. 628-640
Author(s):  
D. E. Andersen ◽  
Peter Kovacic
Keyword(s):  
Molecular Weight ◽  
Tensile Strength ◽  
Active Sites ◽  
Film Structure ◽  
Polymer Chains ◽  
Active Chlorine ◽  
Cross Linking ◽  
Curing Conditions ◽  
Cross Links ◽  
Hydrolysis Of

Abstract Neoprene polymer chains are cross-linked between active chlorine sites on the chains. There are two types of active sites of different reactivity. It is necessary to use more vigorous curing conditions to actuate the less reactive chlorine. The polymer develops cross-links while aging as a latex as well as during cure, and the decline of tensile strength with latex age may be inversely correlated with the amount of cross-linking during aging. The decline of tensile strength can be prevented by compounding with bis (diethylthiocarbamyl) disulfide. The calculations of the swelling data are not so complete as might be desired, primarily because of difficulty of measuring and interpreting the molecular weight of high-conversion polymers and to some uncertainty in determining μ. It is felt that the qualitative observations are justified, and that they serve to define both the technically important effect of film structure on tensile strength and the mechanism of cure.

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