Task 3 8 December 2020
DISTANCE CALCULATION FROM FRET DATA
The protein human serum albumin (HSA) contains
a single tryptophan residue at position 214. HSA
was labeled with b ay attaching anthraniloyl to
cysteine 34 (Fig.1). Normalized emission spectra of
the labeled and unlabeled HSA are shown in Fig. 2.
The Förster distance R0 for FRET pair Trp to
anthraniloyl transfer is listed in Table 2 by your
name. Use the values from Table 1 derived from
emission spectra in Fig. 2 to calculate:
Table 1
1. What is the transfer efficiency E between Trp 214 and anthraniloyl E for
modified HSA? Calculate the E value to two decimal places.
2. What is the distance Trp 214 and anthraniloyl cysteine 34 calculated based on
fluorescence resonance energy transfer?
Indicate the distance value in Angstroms (Å) with precision to one decimaal place.
Send me your answer via email. Correct answer = 1 point.
relative fluorescence
λ (340nm) λ (408nm)
HSA 0.546 0.091
mod HSA 0.202 0.687
Fig. 2
Fig. 1
Table 2
Rcryst R0
1 Dikunová Alžbeta 31.7 29.0
2 Dzurov Matej 33.2 30.4
3 Faturová Jana 31.7 29.0
4 Gašparik Norbert 33.6 30.7
5 Hesko Ondrej 34.5 31.6
6 Jahodová Kateřina 34.0 31.1
7 Kameniarová Michaela 32.6 29.8
8 Konečná Kateřina 30.2 27.6
9 Korytářová Anna 32.6 29.8
10 Kozeleková Aneta 34.6 31.7
11 Kubinyiová Lenka 33.2 30.4
12 Kůřilová Eliška 35.2 32.2
13 Lysáková Klára 33.7 30.8
14 Mikšátková Barbora 33.3 30.5
15 Nováková Barbora 30.7 28.1
16
Prabhullachandran
Unnikannan
35.1 32.1
17 Procházková Markéta 31.1 28.5
18 Šimek Jan 31.8 29.1
19 Tužinčin Dávid 34.1 31.2
This task was prepared base on Problem 1.6 on page 25 in Principles of fluorescence
spectroscopy (2006) book of Prof. Lakowicz.