A stable three-enzyme creatinine biosensor. 1. Impact of structure, function and environment on PEGylated and immobilized sarcosine oxidase

Berberich, Jason A.; Yang, Lee Wei; Madura, Jeffry D.; Bahar, İvet; Russell, Alan J.

doi:10.1016/j.actbio.2004.11.006

Cited by 22 publications

(19 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Also, the use of cellulose acetate cover membranes to prevent silver from reaching the enzyme was studied. Sensors prepared with cover membranes have half‐lives almost an order of magnitude greater than those prepared with no cover membrane over the silver electrode .

{Creatinine + H}_{2 O} \overset{Creatinine amidohydrolase}{} creatine

{Creatinine + H}_{2 O} \overset{Creatine amidinohydrolase}{} sarcosine + urea

\begin{matrix} left Sarcosine + O_{2} + H_{2} normalO \overset{Sarcosine oxidase}{} glycine \\ left + formaldehyde + H_{2} O_{2} \end{matrix}

…”

Section: Biosensors For Important Markers Analysismentioning

confidence: 99%

Application of Electrochemical Biosensors in Clinical Diagnosis

Monošík

Stredansky

Šturdı́k

2012

Clinical Laboratory Analysis

View full text Add to dashboard Cite

Analyses in the clinical area need quick and reliable analytical methods and devices. For this purpose, biosensors can be a suitable option, whereas they are constructed to be simple for use, specific for the target analyte, capable of continuous monitoring and giving quick results, potentially low-costing and portable. In this article, we describe electrochemical biosensors developed for clinical diagnosis, namely for glucose, lactate, cholesterol, urea, creatinine, DNA, antigens, antibodies, and cancer markers assays. Chosen biosensors showed desirable sensitivity, selectivity, and potential for application on real samples. They are often designed to avoid interference with undesired components present in the monitored systems.

show abstract

{Creatinine + H}_{2 O} \overset{Creatinine amidohydrolase}{} creatine

{Creatinine + H}_{2 O} \overset{Creatine amidinohydrolase}{} sarcosine + urea

\begin{matrix} left Sarcosine + O_{2} + H_{2} normalO \overset{Sarcosine oxidase}{} glycine \\ left + formaldehyde + H_{2} O_{2} \end{matrix}

…”

Section: Biosensors For Important Markers Analysismentioning

confidence: 99%

Application of Electrochemical Biosensors in Clinical Diagnosis

Monošík

Stredansky

Šturdı́k

2012

Clinical Laboratory Analysis

View full text Add to dashboard Cite

show abstract

“…Because the site of modification cannot be controlled, the sites that are critical for protein function may be partially or completely blocked upon modification, impacting protein activity. 16 Additionally, the activity of the protein may be further altered by the modification of residues that are involved in protein dynamics (i.e., hinge motions) as well as by the disruption of protein structure. Although natural residues, including cysteines or lysines, may be introduced at specific sites for modification, nonspecific conjugation may still occur if such residues at other sites are not mutated or removed.…”

Section: ■ Introductionmentioning

confidence: 99%

Multisite Clickable Modification of Proteins Using Lipoic Acid Ligase

et al. 2015

View full text Add to dashboard Cite

Approaches that allow bioorthogonal and, in turn, site-specific chemical modification of proteins present considerable opportunities for modulating protein activity and stability. However, the development of such approaches that enable site-selective modification of proteins at multiple positions, including internal sites within a protein, has remained elusive. To overcome this void, we have developed an enzymatic approach for multisite clickable modification based on the incorporation of azide moieties in proteins using lipoic acid ligase (LplA). The ligation of azide moieties to the model protein, green fluorescent protein (GFP), at the N-terminus and two internal sites using lipoic acid ligase was shown to proceed efficiently with near-complete conversion. Modification of the ligated azide groups with poly(ethylene glycol) (PEG), α-d-mannopyranoside, and palmitic acid resulted in highly homogeneous populations of protein-polymer, protein-sugar, and protein-fatty acid conjugates. The homogeneity of the conjugates was confirmed by mass spectrometry (MALDI-TOF) and SDS-PAGE electrophoresis. In the case of PEG attachment, which involved the use of strain-promoted azide-alkyne click chemistry, the conjugation reaction resulted in highly homogeneous PEG-GFP conjugates in less than 30 min. As further demonstration of the utility of this approach, ligated GFP was also covalently immobilized on alkyne-terminated self-assembled monolayers. These results underscore the potential of this approach for, among other applications, site-specific multipoint protein PEGylation, glycosylation, fatty acid modification, and protein immobilization.

show abstract

“…MSOX is a 44 kDa two-domain flavoprotein that catalyzes the oxidative demethylation of sarcosine (N-methyglycine) and is used in the clinical evaluation of renal function and muscle damage (10–15). MSOX is a prototypical member of a family of amino acid oxidases that contain covalently bound FAD (16–19).…”

mentioning

confidence: 99%

Probing Oxygen Activation Sites in Two Flavoprotein Oxidases Using Chloride as an Oxygen Surrogate

et al. 2011

View full text Add to dashboard Cite

A single basic residue above the si-face of the flavin ring is the site of oxygen activation in glucose oxidase (GOX) (His516) and monomeric sarcosine oxidase (MSOX) (Lys265). Crystal structures of both flavoenzymes exhibit a small pocket at the oxygen activation site that might provide a pre-organized binding site for superoxide anion, an obligatory intermediate in the 2-electron reduction of oxygen. Chloride binds at these polar oxygen activation sites, as judged by solution and structural studies. Firstly, chloride forms spectrally detectable complexes with GOX and MSOX. The protonated form of His516 is required for tight binding of chloride to oxidized GOX and for rapid reaction of reduced GOX with oxygen. Formation of a binary MSOX•chloride complex requires Lys265 and is not observed with Lys265Met. Binding of chloride to MSOX does not affect the binding of a sarcosine analog (MTA, methylthioactetate) above the re-face of the flavin ring. Definitive evidence is provided by crystal structures determined for a binary MSOX•chloride complex and a ternary MSOX•chloride•MTA complex. Chloride binds in the small pocket at a position otherwise occupied by a water molecule, and forms hydrogen bonds to four ligands that are arranged in approximate tetrahedral geometry: Lys265:NZ, Arg49:NH1 and two water molecules, one of which is hydrogen bonded to FAD:N5. The results show that chloride: (i) acts as an oxygen surrogate; (ii) is an effective probe of polar oxygen activation sites; (iii) provides a valuable complementary tool to the xenon gas method that is used to map nonpolar oxygen-binding cavities.

show abstract

A stable three-enzyme creatinine biosensor. 1. Impact of structure, function and environment on PEGylated and immobilized sarcosine oxidase

Cited by 22 publications

References 39 publications

Application of Electrochemical Biosensors in Clinical Diagnosis

Application of Electrochemical Biosensors in Clinical Diagnosis

Multisite Clickable Modification of Proteins Using Lipoic Acid Ligase

Probing Oxygen Activation Sites in Two Flavoprotein Oxidases Using Chloride as an Oxygen Surrogate

Contact Info

Product

Resources

About