Research Article Computational Biophysical, Biochemical, and Evolutionary Signature of Human R-Spondin Family Proteins, the Member of Canonical Wnt/-Catenin Signaling Pathway Ashish Ranjan Sharma, 1,2 Chiranjib Chakraborty, 1,3 Sang-Soo Lee, 1 Garima Sharma, 1 Jeong Kyo Yoon, 4 C. George Priya Doss, 5 Dong-Keun Song, 1 and Ju-Suk Nam 1 1 Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Republic of Korea 2 Institute for Skeletal Aging & Orthopedic Surgery, Hallym University Hospital, College of Medicine, Chuncheon-si, Gangwon-do 200-704, Republic of Korea 3 Department of Bioinformatics, School of Computer Sciences, Galgotias University, Greater Noida 203201, India 4 Center for Molecular Medicine, Maine Medial Center Research Institute, 81 Research Drive, Scarborough, ME 04074, USA 5 Medical Biotechnology Division, School of Biosciences and Technology, VIT University, Vellore, Tamil Nadu 632014, India Correspondence should be addressed to Chiranjib Chakraborty; drchiranjib@yahoo.com and Ju-Suk Nam; jsnam88@hallym.ac.kr Received 4 April 2014; Revised 12 July 2014; Accepted 12 July 2014; Published 8 September 2014 Academic Editor: Tun-Wen Pai Copyright © 2014 Ashish Ranjan Sharma et al. his is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In human, Wnt/-catenin signaling pathway plays a signiicant role in cell growth, cell development, and disease pathogenesis. Four human (Rspo)s are known to activate canonical Wnt/-catenin signaling pathway. Presently, (Rspo)s serve as therapeutic target for several human diseases. Henceforth, basic understanding about the molecular properties of (Rspo)s is essential. We approached this issue by interpreting the biochemical and biophysical properties along with molecular evolution of (Rspo)s thorough computational algorithm methods. Our analysis shows that signal peptide length is roughly similar in (Rspo)s family along with similarity in aa distribution pattern. In Rspo3, four N-glycosylation sites were noted. All members are hydrophilic in nature and showed alike GRAVY values, approximately. Conversely, Rspo3 contains the maximum positively charged residues while Rspo4 includes the lowest. Four highly aligned blocks were recorded through Gblocks. Phylogenetic analysis shows Rspo4 is being rooted with Rspo2 and similarly Rspo3 and Rspo1 have the common point of origin. hrough phylogenomics study, we developed a phylogenetic tree of sixty proteins ( = 60) with the orthologs and paralogs seed sequences. Protein-protein network was also illustrated. Results demonstrated in our study may help the future researchers to unfold signiicant physiological and therapeutic properties of (Rspo)s in various disease models. 1. Introduction R-spondins (Rspo)s are a recently discovered family of genes that encodes cysteine-rich secretory proteins containing a thrombospondin type 1 domain/repeat-1 [1]. he (Rspo)s family includes four conserved proteins (Rspo1, Rspo2, Rspo3, and Rspo4), showing overall similarity of 40–60% sequence homology and domain organization [2]. Besides the existence of TSR-1 domain, all four (Rspo)s can be recognized by the existence of a carboxy-terminal region with positively charged amino acids and two furin-like cysteine- rich repeats adjacent to the amino terminus of the mature protein. Numerous studies have implicated (Rspo)s for acting synergistically with extracellular components of the Wnt signaling pathway (Figure 1)[3–5]. Studies showed close or overlapped gene expression of Wnt and (Rspo)s during developmental events, implying a possible coupling of the (Rspo)s with Wnt signaling [6–8]. Consistent with this, a signiicant reduction in mRNA expression of Rspo1 was observed in a Wnt1/3a double knockout mouse [1]. Rspo1 Hindawi Publishing Corporation BioMed Research International Volume 2014, Article ID 974316, 22 pages http://dx.doi.org/10.1155/2014/974316