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Protein
l cc hp cht cao phn t bao gm ch yu l cc L-axit amin kt hp vi nhau qua lin kt peptit
Vai tr:
Xc tc Vn ti Chuyn ng Bo v
Protein
2. Axit amin
Khi nim:
R H2N C H
COOH
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Protein
2. Axit amin
Tn vit tt:
Cu & que
Ho lp th
:N
:H
:O
:C
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(5)
Alanin c hydroxyl ho
Tm bt i
OH + ng =Glycoprotein OH + P = c ch, iu ho th 2
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Cysteine
Serine c nhm sulfhydryl, hay thiol (-SH) thay th cho nhm hydroxyl (-OH)
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Glutamine (Gln, Q)
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Nhm III:
Glutamate hoc axit glutamic(E, Glu) v Aspartat hoc axit aspartic (D, Asp) Lun tch in m pH sinh l(pH 7,4)
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Nhm Iv:
Nhn mt proton pH sinh l (tch in dng) His b ion ha gn pH sinh l; do c th hot ng nh l mt cht nhn proton hoc cho proton ph thuc vo pH ca mi trng xung quanh. (His l mt a.a c tnh baz yu)
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Arginin v Histidin l cc a.a khng thay th tr nh v mt s ngi mc cc bnh ri lon trao i cht (extreme metabolic stress disease)- y l cc axit amin khng thay th c iu kin.
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Ch tr a.a n gin nht l glyxin cn tt c cc axit amin u c cha cacbon bt i, nn chng c tnh quang hot. Hng quay ca mt phng nh sng phn cc c k hiu bng du (+): quay phi v (-): quay tri.
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(2)
Tng in tch: +1
-1
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SPhm thu c (DNP-aa) c mu khng b thy phn trong mi trng axit. Tnh cht ny l c s ca phng php Sanger, mt phng php dng xc nh trinh t a.a trong protein (trinh bay trong phan tinh chat cua
protein).
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Nh cc amin bc nht khc, axit amin p.ng vi axit nitr gii phng kh nit v oxy axit tng ng c hnh thnh:
R H C - NH2 COOH + HO-N=O R H C - OH COOH + N2 + H2 O
Da vo lng nit thot ra ta c th tnh c lng axit amin c trong dung dch (phng php Van Slyke).
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H2N
H+
pH 5-7, toC
COOH
- phn t ninhydrin kh cng vi 1 phn t ninhydrin mi, p. vi NH3b loi ra trn to thnh 1 hp cht c mu tm :
+ NH3+
H
NH4
+ 3 H2 O
+ NH3
R H2N C H
COOH
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Protein
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c gi vng nh lin kt hydro hnh thnh gia nhm CO ca on mch polipeptide ny vi nhm NH ca on mch polypeptide gn k.
(A)
(B)
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:N
:H
:O
:C
:R
Cu trc bc ba
74
V nh hnh
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Kch thc
Ht nano: 1-100 nm; 1 kDa = 1000 Dalton
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4.3. Cc phn ng mu
Phn ng Biure, Foline
Phn ng Folin-Ciocalteu
Giai on 1: Phn ng Biure, 10 pht-RT Giai on 2: Phn ng gia Cu+/axit amin thm (Tyr, Trp) vi thuc th Folin- Ciocalteu (dung dch axit phosphomolybdic-phosphotungstic) ? 30 pht, mu xanh, 750 nm, 10 g/ml
http://www.life.illinois.edu/biochem/355/articles/LowryJBC193_265.pdf http://web.itu.edu.tr/~dulekgurgen/Proteins.pdf
http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/6/47641dat.Par.0001.File.tmp/47641dat.pdf
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1 - 1500g/mL www.freepatentsonline.com/6558717.html
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Luc trng
Kem trng
u ph
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phi by cc lin kt peptide ng vi vng c hiu ca protease, protein bin tnh d tiu ho.
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Protein G vi lp v nc dy 5 A
http://binf.gmu.edu/~greck/Server/intro.php
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Ha tan v Kt ta (tip)
Di tc dng ca cc yu t vt l, ho hc lm trung ho in tch ca cc phn t protein, ph v mng nc, phn t protein s kt hp vi nhau to thnh khi ln, tch khi dung dch kt ta.
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103
http://news.socbay.com/nuoc_cham_o_tphcm_kem_chat_luong_mat_an_toan_-600244572-268632064.html
4.7. To gel, to mng: Khi protein b bin tnh, tp hp li hnh thnh cu trc mng li khng gian 3 chiu c trt t gel protein. Nt cc mng l lin kt a bo hnh thnh gia cc gc R.
www.chembio.uoguelph.ca/.../page6.shtml
104
Lm bn nh tng, to bt
Nh tng=h phn tn ca 2 cht lng ko trn ln, 1 cht dng git nh (0,1-1 mcm) ca pha phn tn, cn cht kia pha lin tc Bt=h phn tn ca cht kh trong pha lin tc l cht lng hoc bn rn
Protein hp ph trn cc b mt, lm bn nh tng v bt theo cc c ch khc nhau Protein thng b bin tnh
Tng tc tnh in
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