Lactoferrin

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Lactoferrin

• Bibi Haniff Thanh Nguyen Ngoc Tran
  Ciamaruel Fears

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Lactoferrin

• What is Lactoferrin?
• Iron-binding protein
• Transports Iron
• Transferrin family
• Possibly Regulates and Maintains Iron levels

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Lactoferrin...

• Moonlighting protein Multifunctional
• 3-D structure
• Some Unknown Functions

Baker,BakerKiddLactoferrin and
transferrinFunctional variations on a common
structural framework,2002
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Lactoferrin

• Locations
• Milk
• Also found in
• Tears
• GI fluid
• Spleen
• Bone
• Brain Tissues

BakerBaker Molecular structure, binding
properties and dynamics of Lactoferrin,2005
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Lactoferrin

• Bovine Lactoferrin (1BLF)
• Species
• Cow/water buffalo
• Human/chimpanzee
• Sheep/goat
• Rats/Mice
• Dog
• Camel
• Pig

BakerBaker Molecular structure, binding
properties and dynamics of Lactoferrin,2005
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Lactoferrin

• Uses
• Multifunctional
• Antimicrobial
• Anti-inflammatory
• Possible Anticancer and regulation of Immune
  system
• Enzymatic Activities
• Cleavage of synthetic substrates

Brock,Jeremy, The physiology of lactoferrin,2002
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Lactoferrin

• Real World Applications
• Infant Formula
• Immune Enhancers
• Prevent/Decrease Osteoporosis

www.google.com/images
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Lactoferrin

• More Real World Applications
• Cure microbial infections
• Injection or Orally
• On-going studies
• Treating diarrhea in
• toddlers
• Other studies majority
• Human LF

www.google.com/images
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Mechanism
Baker,BakerKiddLactoferrin and
transferrinFunctional variations on a common
structural framework,2002
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Structure
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Lactoferrin Structure (1BLF)

• Transferrin family
• X-ray crystallography
• Chain A- AA
• 2 Lobes, 4 Domains

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Structure of Lactoferrin

• 26 Helices
• 27 Beta Strands
• 17 Disulfide bridges

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Ligands

• IRON (Fe3)
• CARBONATE ION (CO3)
• N-ACETYL-D-GLUCOSAMINE
• (NAG )
• BETA-D-MANNOSE
• ALPHA-D-MANNOSE

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FE Binding Sites

• Fe3 binding site 1
• Asp60A
• Tyr92A
• Tyr192A
• His253A
• Fe3 binding site 2
• Asp395A
• Tyr433A
• Tyr526A
• His595A

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AA STATS
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Transmembrane Protein?
- GREASE
-TMHMM
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Motifs of Interest...

• N-glycosylation site
• cAMP- and cGMP-dependent protein kinase
  phosphorylation site Protein
• Kinase C phosphorylation site
• Casein kinase II phosphorylation site
• N-myristoylation site
• Transferrins signatures

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Transferrin Signatures...

• C-lobe
• Transferrin 1
• 11 aa 433-443
• Transferrin 2
• 18 aa 526-543
• Transferrin 3
• 32 aa 568-599
• N-lobe
• Transferrin 1
• 11 aa 92-102
• Transferrin 2
• 18 aa 192-209
• Transferrin 3
• 32 aa 226-257

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Repetitive Structures...

• AVAVVKK
• 94-100 435-441
• KSCHT
• 113-117 455-459
• GYSGAFKCL
• 191-199
• GYTGAFRCL
• 525-533
• GDVAFVKETT V FEN
• 204-217
• GDVAFVKNDTVWEN
• 538-551

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How was my prediction?

• Protein Explorer
• 0 Turns
• 27 Strands
• 26 Helices

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Phylogeny
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Species Used...

• 11 species selected
• All mammalian
• Low E-values
• High score
• High percent identities
• High percent positives

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Phylogeny

• BOXSHADE
• Gene comparison among species
• Highly conserved regions

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CLUSTALW Rooted Tree

• Evolutionary trends
• Human/Chimpanzee
• Water Buffalo/Cow
• Sheep/Goat
• Rodents

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Comparing Trees...

• Show same trends

CLUSTALW Rooted Tree
CLUSTALW Unrooted Tree
Drawtree Unrooted Tree
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Amino Acid vs. Nucleic Acid Tree

• Trends remain the same

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PROTPARS Parsimony Tree

• ONLY 1 Parsimony Tree
• Similar to CLUSTALW UPGMA
• Mouse/Rat
• Human/Chimpanzee
• Cow/Water Buffalo
• Main difference
• Sheep Lt not closely related to Goat

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BOOTSTRAP Tree
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Mutations among species

• Two types of stop codon mutations
• Mutation among 3 stop codons
• TAA to TAG
• TGA to TAA/TAG
• TAA to GAA

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Mutations

• Sequence length variations amongst species
• Mutation of TAA to GAA results in 66bp elongation
  (conserved in R. norvegicus)
• 81bp deletion found in chimpanzee

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Conserved Sequences

• FE1 Binding Site

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FE1 Binding Site

• Mutations affect iron binding
• Favored by natural selection
• Haemophilus influenzae

BakerBaker Molecular structure, binding
properties and dynamics of Lactoferrin,2005
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Work Cited

• Oztas, Yesim,Nuriman Ozgunes. "LactoferrinA
  multifunctional protein." Advances in Molecular
  Medicine 1(2005) 149-154.
• Baker,, EN,Baker,HM. "Molecular Structure,binding
  properties and dynamics of lactoferrin." Cellular
  and Molecular Cell Sciences 62(2005) 2531-2539.
• Roseanu, A,Brock, JH. "What are the Structure and
  Biological Function of Lactoferrin in Human
  Breast Milk?." Life 58(2006) 235-237.
• Kang,, JF,Li, XL,Zhou,RY,Li,LH,Feng,FJ,Guo,XL.
  "Bioinformatics Analysis of Lactoferrin Gene for
  Several Species." Biochem Genet 46(2008)
  312-322.
• Steijns,, JM.,Hooijdonk,ACM.. "Ocurrence,structure
  ,biochemical properties and technological
  characteristics of lactoferrin." British Journal
  of Nutrition 84(2000) S11-S17.
•  Brock,, Jeremy. "The physiology of lactoferrin."
  Biochem 80(2002)
• Baker,, EN,Baker,HM., Kidd, R.. "lactoferrin and
  transferrinFunctional variations on a common
  structural framework." Biochem 80(2002) 27-34.
• http//controlled-trials.com/mrct/trial/407437/bov
  inelactoferrin, accessed November 24th ,2008
• http//www.lactoferrinresearch.org/ accessed
  November 24th, 2008

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Questions?