Characterization of a cysteine-containing peptide tether immobilized onto a gold surface

被引:38
作者
Baas, T
Gamble, L
Hauch, KD
Castner, DG
Sasaki, T
机构
[1] Univ Washington, Dept Chem Engn, Seattle, WA 98195 USA
[2] Univ Washington, Dept Chem, Seattle, WA 98195 USA
[3] Univ Washington, Dept Bioengn, Seattle, WA 98195 USA
关键词
D O I
10.1021/la011847r
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
One side of a triangular molecular tile has been successfully synthesized. The synthesized peptide contains two cysteines (Cys), two glutamic acids (Glu), and a p-aminomethylbenzoyl-p-benzoic acid (AMBBA). The cysteines anchor the peptide to a gold surface by formation of gold-thiolate bonds. The negatively charged glutamic acids should assist in the binding of positively charged proteins to the peptide. The rigid AMBBA provides a benzophenone group for photochemically tethering proteins to the peptide and also acts as the corner for the triangle. Electron spectroscopy for chemical analysis (ESCA) and near-edge X-ray absorption fine structure were used to characterize the composition and orientation of the AMBBA-Cys-Glu-Cys-Glu peptide on the gold surface. Adsorption from 7 muM solutions of the peptide in phosphate-buffered saline produced peptide overlayers with the thiol groups from both cysteines bound to the gold surface. The amide groups of the peptide backbone and the aromatic rings of the AMBBA groups were both oriented parallel to the gold surface. ESCA and fluorescence microscopy demonstrated that albumin could be successfully photoimmobilized to the adsorbed peptide tether. The AMBBA-Cys-Glu-Cys-Glu peptide on gold also could photochemically tether chymotrypsin. The tethered chymotrypsin was active and capable of cleaving a fluorogenic substrate. When the chymotrypsin was tethered as a positively charged protein, it exhibited higher enzyme activity than when it was tethered as a negatively charged protein. However, in all cases the activity of the bound chymotrypsin was lower than its activity in solution.
引用
收藏
页码:4898 / 4902
页数:5
相关论文
共 24 条
  • [11] Micropatterning gradients and controlling surface densities of photoactivatable biomolecules on self-assembled monolayers of oligo(ethylene glycol) alkanethiolates
    Herbert, CB
    McLernon, TL
    Hypolite, CL
    Adams, DN
    Pikus, L
    Huang, CC
    Fields, GB
    Letourneau, PC
    Distefano, MD
    Hu, WS
    [J]. CHEMISTRY & BIOLOGY, 1997, 4 (10): : 731 - 737
  • [12] HUMPHRIES MJ, 1990, J CELL SCI, V97, P585
  • [13] Formation of microscale gradients of protein using heterobifunctional photolinkers
    Hypolite, CL
    McLernon, TL
    Adams, DN
    Chapman, KE
    Herbert, CB
    Huang, CC
    Distefano, MD
    Hu, WS
    [J]. BIOCONJUGATE CHEMISTRY, 1997, 8 (05) : 658 - 663
  • [14] POLYMER METAL INTERFACES STUDIED BY CARBON NEAR-EDGE X-RAY ABSORPTION FINE-STRUCTURE SPECTROSCOPY
    JORDANSWEET, JL
    KOVAC, CA
    GOLDBERG, MJ
    MORAR, JF
    [J]. JOURNAL OF CHEMICAL PHYSICS, 1988, 89 (04) : 2482 - 2489
  • [15] Structures and dynamic formation processes of porphyrin adlayers on iodine-modified Au(111) in solution: In situ STM study
    Kunitake, M
    Akiba, U
    Batina, N
    Itaya, K
    [J]. LANGMUIR, 1997, 13 (06) : 1607 - 1615
  • [16] SELF-ORGANIZED PORPHYRIN ARRAY ON IODINE-MODIFIED AU(111) IN ELECTROLYTE-SOLUTIONS - IN-SITU SCANNING-TUNNELING-MICROSCOPY STUDY
    KUNITAKE, M
    BATINA, N
    ITAYA, K
    [J]. LANGMUIR, 1995, 11 (07) : 2337 - 2340
  • [17] Maheshwari G, 2000, J CELL SCI, V113, P1677
  • [18] AN RGD SPACING OF 440NM IS SUFFICIENT FOR INTEGRIN ALPHA-V-BETA-3-MEDIATED FIBROBLAST SPREADING AND 140NM FOR FOCAL CONTACT AND STRESS FIBER FORMATION
    MASSIA, SP
    HUBBELL, JA
    [J]. JOURNAL OF CELL BIOLOGY, 1991, 114 (05) : 1089 - 1100
  • [19] Tailored substrates for studies of attached cell culture
    Mrksich, M
    [J]. CELLULAR AND MOLECULAR LIFE SCIENCES, 1998, 54 (07) : 653 - 662
  • [20] Surface characterization of mixed self-assembled monolayers designed for streptavidin immobilization
    Nelson, KE
    Gamble, L
    Jung, LS
    Boeckl, MS
    Naeemi, E
    Golledge, SL
    Sasaki, T
    Castner, DG
    Campbell, CT
    Stayton, PS
    [J]. LANGMUIR, 2001, 17 (09) : 2807 - 2816