Nilapwar Sanjay M, Nardelli Maria, Westerhoff Hans V, Verma Malkhey
Manchester Interdisciplinary Biocentre, The University of Manchester, Manchester, United Kingdom.
Methods Enzymol. 2011;500:59-75. doi: 10.1016/B978-0-12-385118-5.00004-9.
Absorption spectroscopy is one of the most widely used techniques employed for determining the concentrations of absorbing species (chromophores) in solutions. It is a nondestructive technique which biologists and biochemists and now systems biologists use to quantify the cellular components and characteristic parameters of functional molecules. This quantification is most relevant in the context of systems biology. For creating a quantitative depiction of a metabolic pathway, a number of parameters and variables are important and these need to be determined experimentally. This chapter describes the UV-visible absorption spectroscopy used to produce experimental data for bottom-up modeling approaches of systems biology which uses concentrations and kinetic parameters (K(m) and V(max)) of enzymes of metabolic/signaling pathways, intracellular concentrations of metabolites and fluxes. It also briefly describes the application of this technique for quantification of biomolecules and investigating biomolecular interactions.
吸收光谱法是用于测定溶液中吸收物种（发色团）浓度的最广泛使用的技术之一。它是一种无损技术，生物学家、生物化学家以及现在的系统生物学家都用它来量化细胞成分和功能分子的特征参数。这种量化在系统生物学背景下最为相关。为了创建代谢途径的定量描述，许多参数和变量很重要，这些需要通过实验来确定。本章描述了紫外可见吸收光谱法，该方法用于为系统生物学的自下而上建模方法生成实验数据，这些方法使用代谢/信号通路中酶的浓度和动力学参数（K(m)和V(max)）、代谢物的细胞内浓度和通量。它还简要描述了该技术在生物分子定量和研究生物分子相互作用方面的应用。
