Protein concentration is not an absolute prerequisite for the determination of secondary structure from circular dichroism spectra: a new scaling method

Bibliographic Details
Title: Protein concentration is not an absolute prerequisite for the determination of secondary structure from circular dichroism spectra: a new scaling method
Authors: Raussens, Vincent, Ruysschaert, Jean Marie, Goormaghtigh, Erik
Source: Analytical Biochemistry. 319:114-121
Publisher Information: Elsevier BV, 2003.
Publication Year: 2003
Subject Terms: 0301 basic medicine, Protein Structure, Secondary, 0303 health sciences, 03 medical and health sciences, Circular Dichroism -- methods, Circular Dichroism, Proteins -- pharmacology, Chimie, Proteins, Proteins -- chemistry, Sensitivity and Specificity, Protein Structure, Secondary
Description: We present here a simple and rapid method to extract good estimates of protein secondary structure content from circular dichroism (CD) spectra without any prior knowledge of the sample concentration. The method involves two steps: first, a single-wavelength normalization procedure and, second, the application for each secondary structure of a quadratic model based on one or two wavelength intensities. These quadratic models were derived by a cross-validation analysis of a new protein CD spectrum database. Tested on CD spectra of proteins at different concentrations, the normalization was shown to render the method virtually independent of the sample concentration. Further tests on CD spectra not recorded in our laboratory showed that our quadratic models are of general applicability. Even though the success of the present approach is less than that for currently available methods, its simplicity and the fact that the concentration is not needed may be very attractive for the study of small amounts of membrane proteins or peptides for which an accurate concentration determination might be very difficult or impossible to obtain.
Document Type: Article
File Description: 1 full-text file(s): application/pdf
Language: English
ISSN: 0003-2697
DOI: 10.1016/s0003-2697(03)00285-9
Access URL: https://pubmed.ncbi.nlm.nih.gov/12842114
https://www.sciencedirect.com/science/article/pii/S0003269703002859
https://www.ncbi.nlm.nih.gov/pubmed/12842114
https://difusion.ulb.ac.be/vufind/Record/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/77591/Details
https://europepmc.org/article/MED/12842114
Rights: Elsevier TDM
Accession Number: edsair.doi.dedup.....88f90c7e07aa2e43a556f87c1b16e3dd
Database: OpenAIRE
Description
ISSN:00032697
DOI:10.1016/s0003-2697(03)00285-9