When you immobilized enzyme on polymeric support can you detect changes in protein content (αhelix% or β sheet% in enzyme)?
Yes, it has been done and you can measure the changes in secondary structure via CD. Have a look at this paper:
Secondary structure of proteins associated in thin films.
Safar J, Roller PP, Ruben GC, Gajdusek DC, Gibbs CJ Jr.
Biopolymers. 1993 Sep;33(9):1461-76.
Yes, it has been done and you can measure the changes in secondary structure via CD. Have a look at this paper:
Secondary structure of proteins associated in thin films.
Safar J, Roller PP, Ruben GC, Gajdusek DC, Gibbs CJ Jr.
Biopolymers. 1993 Sep;33(9):1461-76.
Hi Parastou, as Holger said, there are ways to do CD on immobilized protein if you can make a membrane or a film of the immobilized protein.
But, if you have a Jasco CD instrument with the circular cuvettes, then things become more complicated.
These specs. will only give you a good readout if the protein is soluble and evenly distributed in the cuvette.
IR spectroscopy is probably better than CD for immobilized proteins in a thin film for the reasons Steingrimur suggested.
Additionally to the reference of Holger there are some further ones on CD (UV/VIS region) of protein films or in solid state including correlations to FTIR technique:
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-CIRCULAR DICHROISM OF POLYPEPTIDE AND PROTEIN CONFORMATIONS - FILM STUDIES, FASMAN, GD; HOVING, H; TIMASHEF.SN
BIOCHEMISTRY 1970, 9(17), 3316.
-ORIENTED SECONDARY STRUCTURE IN INTEGRAL MEMBRANE-PROTEINS .1. CIRCULAR-DICHROISM AND INFRARED-SPECTROSCOPY OF CYTOCHROME-OXIDASE IN MULTILAMELLAR FILMS, BAZZI, MD; WOODY, RW, BIOPHYSICAL JOURNAL 1985, 48(6), 957-966
-Protein secondary structure content in solution, films and tissues: Redundancy and complementarity of the information content in circular dichroism, transmission and ATR FTIR spectra
Goormaghtigh, Erik; Gasper, Regis; Benard, Audrey; et al.
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS 2009, 1794 (9), 1332-1343
-Circular dichroism studies on proteins in films and in solution: Estimation of secondary structure by g-factor analysis
McPhie, P, ANALYTICAL BIOCHEMISTRY 2001, 293 (1), 109-119
-Secondary structure analyses of protein films on gold surfaces by circular dichroism
Shimizu, M; Kazutoshi, K; Morii, H; et al.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2003, 310 (2), 606-611.
-Circular dichroism measurement of a protein in dried thin films
By: Harada, T; Kuroda, R
CHEMISTRY LETTERS 2002, 3 326-327.
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But take care. CD spectra may be influenced by the rotation position of the protein film sample. Best take spectra at at least some 4 rotation angles (0, 90, 180, 270°) and compare.
Best regards,
Martin
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