NDLI: Molecular handles for the mechanical manipulation of single-membrane proteins in living cells

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Molecular handles for the mechanical manipulation of single-membrane proteins in living cells

Content Provider	IEEE Xplore Digital Library
Author	Gorostiza, P. Tombola, F. Verdaguer, A. Smith, S.B. Bustamante, C. Isacoff, E.Y.
Copyright Year	2002
Abstract	We have developed a procedure to selectively biotinylate a specific membrane protein, enabling its attachment to external force probes and thus allowing its mechanical manipulation within its native environment. Using potassium channels as model membrane proteins in oocytes, we have found that Maleimide-PEG3400-biotin is the crosslinker with highest conjugation selectivity and accessibility to external probes. Neutravidin-coated beads provide for directed attachment while avoiding nonspecific interactions with the cell. The technology was successfully tested by mechanical manipulation of biotinylated extracellular residues of channels in oocytes using an atomic force microscope under conditions which preserve function of the channels. Binding forces of /spl sim/80 pN at 100 nN/s were measured.
Sponsorship	IEEE Engineering in Medicine and Biology Society IEEE Systems, Man, and Cybernetics Society IEEE Nanotechnology Council IEEE Computational Intelligence Society IEEE Robotics and Automation Society IEEE Computer Society
Page Count	8
File Size	756745
Starting Page	269
Ending Page	276
File Format	PDF
ISSN	15361241
Volume Number	4
Issue Number	4
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2005-12-01
Publisher Place	U.S.A.
Access Restriction	One Nation One Subscription (ONOS)
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Biomembranes Probes Protein engineering Biological cells Force measurement Physics Extracellular Atomic force microscopy DNA Microorganisms single-molecule force measurements Atomic force spectroscopy living cells patch clamp selective molecular attachment
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Medicine Bioengineering Electrical and Electronic Engineering Biomedical Engineering Computer Science Applications Biotechnology Pharmaceutical Science

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