Cysteine-specific surface tethering of genetically engineered cytochromes for fabrication of metalloprotein nanostructures

Journal Article · Sat Jan 01 00:00:00 EST 1994 · Langmuir; (United States)

DOI:https://doi.org/10.1021/la00013a023· OSTI ID:5146819

Hong, H G; Jiang, M; Sligar, S G; Bohn, P W ^[1]

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1. Univ. of Illinois, Urbana-Champaign, IL (United States)

The preparation of oriented metalloprotein nanostructures through introduction of specific and complementary reactive groups on the solid and protein surfaces is critically dependent on the reaction conditions used to prepare the solid surface. Key problems include the hydrolytic stability of the Si-O bond, the low reactivity of simple nucleophilic silane reagents, protein physisorption, and identification of conditions for producing monolayer protein coverages. These problems are largely circumvented by utilizing a two-step linker synthesis, in which the surface is first prepared with a monolayer of (3-aminopropyl)silane (3-APS), and the resulting structure is derivatized with the heterobifunctional reagent N-succinimidyl 6-maleimidocaproate (EMCS). The maleimide functionality is then presented to the protein, into which a single unique cysteine residue has been introduced by genetic engineering techniques. Hydrolytic stability is dramatically enhanced by including a postreaction curing step, in which the solid surface temperature is elevated to drive the alkoxylsilane condensation reaction to completion. Finally substituting a gas-phase chemical vapor deposition procedure for the liquid-phase reaction of the 3-APS produces dramatically better control over coverage and quality of the resulting films. 23 refs., 4 figs., 2 tabs.

• DOE Contract Number: FG02-88ER13949
• OSTI ID: 5146819
• Journal Information: Langmuir; (United States), Vol. 10:1; ISSN 0743-7463
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
CYTOCHROMES
CHEMICAL REACTIONS
METALLOPROTEINS
CHEMICAL VAPOR DEPOSITION
CYSTEINE
DERIVATIZATION
FILMS
GENETIC ENGINEERING
SILANES
SPECTROSCOPY
AMINO ACIDS
BIOTECHNOLOGY
CARBOXYLIC ACIDS
CHEMICAL COATING
DEPOSITION
HYDRIDES
HYDROGEN COMPOUNDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SILICON COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PIGMENTS
PROTEINS
SILICON COMPOUNDS
SURFACE COATING
THIOLS
400201* - Chemical & Physicochemical Properties
400102 - Chemical & Spectral Procedures
550200 - Biochemistry