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Gallinacin 2

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Gallinacin 2 is a cysteine-rich antimicrobial peptide of chicken leukocytes.

Category

Functional Peptides

Catalog number

BAT-012151

CAS number

156409-45-9

Molecular Formula

C205H322N60O48S6

Molecular Weight

4587.50

Specification
Reference Reading

IUPAC Name

(3S)-3-[[(2S)-2-[[(2S)-6-amino-2-[[(2S)-2-[(2-aminoacetyl)amino]-5-carbamimidamidopentanoyl]amino]hexanoyl]amino]-3-hydroxypropanoyl]amino]-4-[[(2R)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-4-amino-1-[[2-[[(2S)-1-[[(2R)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2R)-1-[(2S)-2-[[(2S)-1-[[(2S)-1-[[(2S,3R)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[2-[[(2S)-6-amino-1-[[(2R)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[(2S)-2-[[(2S,3S)-2-[[(2S)-2-[[(2S)-6-amino-2-[[(2R)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-amino-3-(4H-imidazol-4-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-sulfanylpropanoyl]amino]-3-sulfanylpropanoyl]amino]hexanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]oxy-1-oxo-3-phenylpropan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1-oxo-3-sulfanylpropan-2-yl]amino]-1-oxohexan-2-yl]amino]-2-oxoethyl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]carbamoyl]pyrrolidin-1-yl]-1-oxo-3-sulfanylpropan-2-yl]amino]-1-oxohexan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxo-3-sulfanylpropan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-2-oxoethyl]amino]-1,4-dioxobutan-2-yl]amino]-1-oxohexan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-1-oxo-3-sulfanylpropan-2-yl]amino]-4-oxobutanoic acid

Synonyms

Chp2 peptide; Thp2 peptide; gallinacin 2 protein, chicken

Sequence

LFCKGGSCHFGGCPSHLIKVGSCFGFRSCCKWPWNA

InChI

InChI=1S/C205H322N60O48S6/c1-15-112(11)163(262-178(289)135(64-44-78-226-205(220)221)237-172(283)129(57-32-37-71-207)241-191(302)152(102-315)260-194(305)155(105-318)258-180(291)136(80-108(3)4)243-167(278)125(212)90-122-95-222-107-230-122)196(307)252-147(89-121-94-227-126-55-30-29-54-124(121)126)201(312)313-200(311)146(88-119-52-27-20-28-53-119)251-175(286)134(63-43-77-225-204(218)219)240-188(299)149(99-267)254-193(304)154(104-317)256-171(282)127(56-31-36-70-206)233-160(273)96-229-169(280)148(98-266)255-197(308)164(113(12)16-2)263-186(297)139(83-111(9)10)250-198(309)165(115(14)269)264-187(298)138(82-110(7)8)245-184(295)143(87-120-66-68-123(270)69-67-120)249-195(306)157-65-45-79-265(157)199(310)156(106-319)261-177(288)132(60-35-40-74-210)238-179(290)137(81-109(5)6)244-183(294)141(85-117-48-23-18-24-49-117)242-166(277)114(13)231-190(301)151(101-314)257-181(292)140(84-116-46-21-17-22-47-116)234-161(274)97-228-168(279)144(91-158(213)271)247-174(285)130(58-33-38-72-208)236-173(284)133(62-42-76-224-203(216)217)239-182(293)142(86-118-50-25-19-26-51-118)246-192(303)153(103-316)259-185(296)145(92-162(275)276)248-189(300)150(100-268)253-176(287)131(59-34-39-73-209)235-170(281)128(232-159(272)93-211)61-41-75-223-202(214)215/h17-30,46-55,66-69,94-95,107-115,122,125,127-157,163-165,227,266-270,314-319H,15-16,31-45,56-65,70-93,96-106,206-212H2,1-14H3,(H2,213,271)(H,228,279)(H,229,280)(H,231,301)(H,232,272)(H,233,273)(H,234,274)(H,235,281)(H,236,284)(H,237,283)(H,238,290)(H,239,293)(H,240,299)(H,241,302)(H,242,277)(H,243,278)(H,244,294)(H,245,295)(H,246,303)(H,247,285)(H,248,300)(H,249,306)(H,250,309)(H,251,286)(H,252,307)(H,253,287)(H,254,304)(H,255,308)(H,256,282)(H,257,292)(H,258,291)(H,259,296)(H,260,305)(H,261,288)(H,262,289)(H,263,297)(H,264,298)(H,275,276)(H4,214,215,223)(H4,216,217,224)(H4,218,219,225)(H4,220,221,226)/t112-,113-,114-,115+,122?,125-,127-,128-,129-,130-,131-,132-,133-,134-,135-,136-,137-,138-,139-,140-,141-,142-,143-,144-,145-,146-,147-,148-,149-,150-,151-,152-,153-,154-,155-,156-,157-,163-,164-,165-/m0/s1

InChI Key

SGVJXVIMIBPWHX-XALWAODASA-N

Canonical SMILES

CCC(C)C(C(=O)NC(CC1=CNC2=CC=CC=C21)C(=O)OC(=O)C(CC3=CC=CC=C3)NC(=O)C(CCCNC(=N)N)NC(=O)C(CO)NC(=O)C(CS)NC(=O)C(CCCCN)NC(=O)CNC(=O)C(CO)NC(=O)C(C(C)CC)NC(=O)C(CC(C)C)NC(=O)C(C(C)O)NC(=O)C(CC(C)C)NC(=O)C(CC4=CC=C(C=C4)O)NC(=O)C5CCCN5C(=O)C(CS)NC(=O)C(CCCCN)NC(=O)C(CC(C)C)NC(=O)C(CC6=CC=CC=C6)NC(=O)C(C)NC(=O)C(CS)NC(=O)C(CC7=CC=CC=C7)NC(=O)CNC(=O)C(CC(=O)N)NC(=O)C(CCCCN)NC(=O)C(CCCNC(=N)N)NC(=O)C(CC8=CC=CC=C8)NC(=O)C(CS)NC(=O)C(CC(=O)O)NC(=O)C(CO)NC(=O)C(CCCCN)NC(=O)C(CCCNC(=N)N)NC(=O)CN)NC(=O)C(CCCNC(=N)N)NC(=O)C(CCCCN)NC(=O)C(CS)NC(=O)C(CS)NC(=O)C(CC(C)C)NC(=O)C(CC9C=NC=N9)N

1. Polymorphic characterisation of gallinacin candidate genes and their molecular associations with growth and immunity traits in chickens

M S Saleh, M H Khalil, M M Iraqi, A Camarda Br Poult Sci. 2021 Apr;62(2):180-187. doi: 10.1080/00071668.2020.1847252. Epub 2020 Dec 14.

1.Four gallinacin (GAL) genes were assessed by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) to characterise these genes in Fayoumi (F) and Rhode Island Red (R) breeds and their crosses of Rhode Island Red × Fayoumi (½R½F) and Fayoumi × Rhode Island Red (½F½R).2.Genes examined were GAL2, GAL3, GAL4 and GAL5. The molecular associations between the SNPs of the gallinacin genes and body weight, caecal bacterial count and the serum antibody titres of IgA, IgG and IgM were determined. In the R breed, the frequency of TC genotype was higher than TT and CC genotypes for the GAL3 gene. The GG genotype frequency was higher than AA and AG genotypes for the GAL4 gene in the other genetic groups, and the CA genotype frequency was higher than CC and AA genotypes in crosses for the GAL5 gene.3.In all populations, the frequency of the C allele was higher than the T allele for the GAL3 gene, the G allele was higher than the A allele for the GAL4 gene and the C allele was higher than the A allele for the GAL5 gene. The observed heterozygosity in R, ½R½F and ½F½R was 0.476, 0.375 and 0.158 for the GAL3 gene, 0.458, 0.615 and 0.250 for the GAL4 gene and 0.053, 0.792 and 0.739 for the GAL5 gene, while the expected heterozygosities were 0.490, 0.430 and 0.145 for the GAL3 gene, 0.430, 0.348 and 0.219 for the GAL4 gene and 0.229, 0.478 and 0.496 for the GAL5 gene, respectively.4.On a molecular level, the genotype TT was significantly higher for body weight than TC and CC genotypes in the GAL3 gene. Birds with the GG genotype had a significantly lower Salmonella typhimurium count than birds with AA genotype in the GAL4 gene. Birds with the genotype AA had higher significant body weights than those with CC and CA genotypes in the GAL5 gene.5.The results indicated that the GAL3, GAL4 and GAL5 genes are potential candidates for selection programmes to improve S. typhimurium resistance and body weight in chickens.

2. Gallinacin-3, an inducible epithelial beta-defensin in the chicken

C Zhao, T Nguyen, L Liu, R E Sacco, K A Brogden, R I Lehrer Infect Immun. 2001 Apr;69(4):2684-91. doi: 10.1128/IAI.69.4.2684-2691.2001.

Gallinacin-3 and gallopavin-1 (GPV-1) are newly characterized, epithelial beta-defensins of the chicken (Gallus gallus) and turkey (Meleagris gallopavo), respectively. In normal chickens, the expression of gallinacin-3 was especially prominent in the tongue, bursa of Fabricius, and trachea. It also occurred in other organs, including the skin, esophagus, air sacs, large intestine, and kidney. Tracheal expression of gallinacin-3 increased significantly after experimental infection of chickens with Haemophilus paragallinarum, whereas its expression in the tongue, esophagus, and bursa of Fabricius was unaffected. The precursor of gallinacin-3 contained a long C-terminal extension not present in the prepropeptide. By comparing the cDNA sequences of gallinacin-3 and GPV-1, we concluded that a 2-nucleotide insertion into the gallinacin-3 gene had induced a frameshift that read through the original stop codon and allowed the chicken propeptide to lengthen. The striking structural resemblance of the precursors of beta-defensins to those of crotamines (highly toxic peptides found in rattlesnake venom) supports their homology, even though defensins are specialized to kill microorganisms and crotamines are specialized to kill much larger prey.

3. Direct screening identifies mature beta-defensin 2 in avian heterophils

L Kannan, N C Rath, R Liyanage, J O Lay Jr Poult Sci. 2009 Feb;88(2):372-9. doi: 10.3382/ps.2008-00366.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to screen avian heterophils in the m/z range of 1 to 20 kDa with an objective to identify specific cell-associated peptides that may be reflective of their functional physiology. The MALDI-TOF-MS profiles of crude heterophil extract showed a high intensity peak with average mass of m/z 3916.1 for chicken and m/z 4129.6 for turkey. To identify these peaks, we first purified m/z 3916.1 from chicken bone marrow extract using reverse-phase high performance liquid chromatography (RP-HPLC). Edman sequencing and peptide mass fingerprinting exclusively confirmed this peptide as beta-defensin 2 (BD2) or gallinacin-2, a broad-range antimicrobial peptide. A Uniprot database search followed by the MASCOT sequence query revealed m/z 4129.6 to be the corresponding turkey ortholog of avian beta-defensin 2 (AvBD2), also called turkey heterophil peptide 2. Both AvBD2 peptides are 36 amino acids long including a highly conserved region with 6 invariant cysteines forming the 3 disulfide bonds characteristic of defensins. The method confirmed the existence of the complete mature peptide sequence of the turkey heterophilic BD2 previously proposed based on cDNA analysis. These results demonstrate that screening of the crude extract by MALDI-TOF-MS can identify cell- or tissue-associated peptides in their functional or mature forms, raising the possibility that such peptides can be used as biomarkers in their altered physiological states.