Sensitive detection of sulfhydryl groups in surface-confined metallothioneins and related species via ferrocene-capped gold nanoparticle/streptavidin conjugates.
Metallothionein (MT), a cysteine-rich metalloprotein that is purported to play an important role in heavy metal accumulation and detoxification, and its related peptidic species were attached onto dithiobissuccinimidyl propionate self-assembled monolayers. The spatially accessible sulfhydryl groups present in these immobilized biomolecules, tagged with N-biotinoyl-N'-[6-maleimidohexanoyl]hydrazide, were detected voltammetrically at a sensitive level via the use of ferrocene (Fc)-capped gold nanoparticle/streptavidin conjugates. The method was established first by examining relatively simple peptides (e.g., glutathione). For the hexapeptidic species that resembles the N-terminus of MT with a sequence of Lys-Cys-Thr-Cys-Cys-Ala, concentration levels as low as 0.050 nM can be determined. Such a remarkable sensitivity is attributed to the presence of a large number of Fc caps present at each gold nanoparticle, which enhances the detection of a small number of surface-bound sulfhydryl groups. Microgravimetric measurements, performed with a quartz crystal microbalance, were used in tandem with voltammetry to quantify the number of tagged sulfhydryl groups. Through extraction of the metals present in MT adsorbate, it is demonstrated that this amplified voltammetric detection is also suitable for the investigation of the variation of the number of sulfhydryl groups present at an electrode and sensitive to the change of surface structure of an immobilized biomolecule. This work represents a new method for the determination of sulfhydryl groups inherent in surface-bound proteins or peptides and can facilitate the study on the environmental issues related to MTs.
Jimenez
Omar A
OA
Department of Chemistry and Biochemistry, California State University, Los Angeles, Los Angeles, California 90032, USA.
Chikneyan
Sara
S
Baca
Alfred J
AJ
Wang
Jianxiu
J
Zhou
Feimeng
F
D004784:Environmental Monitoring / Q000379:methods*; D006046:Gold / Q000737:chemistry; D008668:Metallothionein / Q000032:analysis* / Q000737:chemistry*; D019216:Metals, Heavy / Q000737:chemistry; D049329:Nanostructures; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000032:analysis*; D014873:Water Pollutants / Q000032:analysis*
D019216:Metals, Heavy; D013438:Sulfhydryl Compounds; D014873:Water Pollutants; D006046:Gold; D019809:Streptavidin; D008668:Metallothionein