title: Investigation and identification of protein γ-glutamyl carboxylation sites.
abstract: BACKGROUND
  Carboxylation is a modification of glutamate (Glu) residues which occurs post-translation that is catalyzed by γ-glutamyl carboxylase in the lumen of the endoplasmic reticulum. Vitamin K is a critical co-factor in the post-translational conversion of Glu residues to γ-carboxyglutamate (Gla) residues. It has been shown that the process of carboxylation is involved in the blood clotting cascade, bone growth, and extraosseous calcification. However, studies in this field have been limited by the difficulty of experimentally studying substrate site specificity in γ-glutamyl carboxylation. In silico investigations have the potential for characterizing carboxylated sites before experiments are carried out.
  
  
  RESULTS
  Because of the importance of γ-glutamyl carboxylation in biological mechanisms, this study investigates the substrate site specificity in carboxylation sites. It considers not only the composition of amino acids that surround carboxylation sites, but also the structural characteristics of these sites, including secondary structure and solvent-accessible surface area (ASA). The explored features are used to establish a predictive model for differentiating between carboxylation sites and non-carboxylation sites. A support vector machine (SVM) is employed to establish a predictive model with various features. A five-fold cross-validation evaluation reveals that the SVM model, trained with the combined features of positional weighted matrix (PWM), amino acid composition (AAC), and ASA, yields the highest accuracy (0.892). Furthermore, an independent testing set is constructed to evaluate whether the predictive model is over-fitted to the training set.
  
  
  CONCLUSIONS
  Independent testing data that did not undergo the cross-validation process shows that the proposed model can differentiate between carboxylation sites and non-carboxylation sites. This investigation is the first to study carboxylation sites and to develop a system for identifying them. The proposed method is a practical means of preliminary analysis and greatly diminishes the total number of potential carboxylation sites requiring further experimental confirmation.
authors.affiliation: Department of Computer Science and Engineering, Yuan Ze University, Chung-Li 320, Taiwan. francis@saturn.yzu.edu.tw
authors.forename: Tzong-Yi
authors.initials: TY
authors.lastname: Lee
authors.forename: Cheng-Tsung
authors.initials: CT
authors.lastname: Lu
authors.forename: Shu-An
authors.initials: SA
authors.lastname: Chen
authors.forename: Neil Arvin
authors.initials: NA
authors.lastname: Bretaña
authors.forename: Tzu-Hsiu
authors.initials: TH
authors.lastname: Cheng
authors.forename: Min-Gang
authors.initials: MG
authors.lastname: Su
authors.forename: Kai-Yao
authors.initials: KY
authors.lastname: Huang
mesh_terms: D015055:1-Carboxyglutamic Acid / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D060388:Support Vector Machine; D014812:Vitamin K / Q000737:chemistry
chemical_list: D011506:Proteins; D014812:Vitamin K; D015055:1-Carboxyglutamic Acid