Accelerated evolution of small serum proteins (SSPs)—The PSP94 family proteins in a Japanese viper

Accelerated Evolution of Fetuin Family Proteins inProtobothrops flavoviridis(Habu Snake) Serum and the Discovery of an L1-Like Genomic Element in the Intronic Sequence of a Fetuin-Encoding Gene

“…As in previous studies involving PLA 2 inhibitor 63) and SSPs, 25) an unusual accumulation of nucleotide substitutions and higher nonsynonymous substitution rates were found in specific coding exons. Thus, in these genes, we found a signature of accelerated evolution similar to the genes for venom enzymes, but we further found a tendency for these substitutions to occur intensively in limited, narrow regions.…”

“…The reaction mixtures were incubated at 25 C, 37 C, 42 C, and 50 C for 1 h, and the resultant cDNA fragments were analyzed on a 6% acrylamide/8 M urea/TBE gel, followed by visualization with an SR2040 imaging plate and a FLA3000 fluorescent image analyzer (Fujifilm, Tokyo).…”

“…6215; Fax: +81-92-865-6030; E-mail: deshi@fukuoka-u.ac.jp Abbreviations: L1, LINE1; HSF, habu serum factor; HLP, HSF-like protein; PfL1, L1 in the habu snake genome; ORF, open reading frame; RT, reverse transcriptase; PfRT, habu snake RT-like protein; PLA 2 , phospholipase A 2 ; MP, metalloprotease; CY, cystatin-like domain; HR, histidine-rich domain; MSF, mamushi serum factor; MLP, MSF-like protein; SSP, small serum protein; K N , nucleotide substitutions per site; K S , synonymous substitutions per site; K A , nonsynonymous substitutions per site; SP, signal peptide; AHSG, 2-HS-glycoprotein lation of nonsynonymous nucleotide substitutions in the protein-coding region encoding small serum proteins (SSPs), as compared to the noncoding region of the cDNA sequences in the habu. 24,25) This indicates that accelerated evolution has also occurred in SSPs, implying that toxic enzymes and compensatory inhibitors co-evolved in a unique evolutionary manner. However, whether accelerated evolution of venomous snake serum protein families is universal, as is the case for venom enzyme families, remains unknown.…”

Structural analysis and characterization of new small serum proteins from the serum of a venomous snake (Gloydius blomhoffii)

“…Full-length cDNA for mSSP-1 to mSSP-4 was obtained by PCR, using a pair of oligonucleotides ssp-sigN that had originally been designed based on the cDNA nucleotide sequence for 5′-UTR, highly conserved between hSSPs (5′-UTR at 12 bp upstream of the start codon 19) ) and an adaptor-linked oligo(dT) primer. Several positive clones of the expected size (approx.…”

“…Each of the full-length cDNAs encode a mature protein and a highly conserved signal peptide. 19) *Corresponding author. Email: anarumi@fukuoka-u.ac.jp Abbreviations: CRISP, cysteine-rich secretory protein; FTC, fluorescein thiocarbamoyl; MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight; MSF, mamushi serum factor; NBS, N-bromosuccinimide; ORF, open reading frame; PAGE, polyacrylamide gel electrophoresis; PCR, polymerase chain reaction; Pe, S-pyridylethylated; PLI, phospholipase A 2 inhibitor; PSP94, prostatic prostate secretory protein of 94 amino acids; RP-HPLC, reverse-phase high-performance liquid chromatography; SSP, small serum protein; SVMP, snake venom metalloproteinase; TNBS, 2,4,6-trinitrobenzenesulphonic acid; UTR, untranslated region.…”